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Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329845
Dataset. 2022

ADDITIONAL FILE 1 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 1: Figure S1. a, Relative mRNA expression of RRAS2 in different types of leukemia. Data comes from (Haferlach et al., 2010) and has been retrieved from www.oncomine.org . b, Schematic representation of the overexpression cassette inserted into the Rosa26 locus. c, Relative expression of RRAS2 measured by RT-qPCR in different organs of Rosa26-RRAS2fl/flxSox2-Cre (Sox2-Cre+) mice compared to that of WT C57BL/6 J Control mice using 18S as the reference gene. All expression numbers were normalized to those of liver from WT Control mice (mean = 1). Data show relative expression of RRAS2 in the indicated organs in n = 3–4 8 month-old independent mice. d, Quantification of spleen weight from control and 6 month-old Sox2-Cre + mice. Data shown correspond to four control mice and eleven Sox2-Cre mice. Two-tailed unpaired t-test with Welch’s correction. e, Two-parameter flow cytometry of the expression of CD5 and IgM in B cells in the spleen of 6 month-old control and Sox2-Cre + mice. f, Quantification of the number of CD5 + IgM+ B cells in the spleens and bone marrow of 6 month-old control and Sox2-Cre + mice. Data correspond to triplicate measurements of one control and three Sox2-Cre mice. Unpaired t-test with Welch’s correction. g, Quantification of the serum IgM concentration in the blood of 35–40 wk-old control (n = 3) and mb1-Cre (n = 8) mice by ELISA. Unpaired t-test with Welch’s correction. h, Representative images from Giemsa stainings of blood smears of 36 wk-old control and mb1-Cre mice. i, Two-parameter flow cytometry of the forward scatter and CD5 expression in CD19+ cells in the blood of 16 wk-old mb1-Cre mice. The gated population represents large cells. j, Two-parameter flow cytometry of CD5 expression and BrdU incorporation in CD19+ cells in the blood of 16 wk-old mb1-Cre mice. k, Quantification of the percentage of CD19+ cells that are CD5+ blasts and of the CD19+ CD5+ cells that have incorporated BrdU., Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329845
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329845
HANDLE: http://hdl.handle.net/10261/329845
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329845
PMID: http://hdl.handle.net/10261/329845
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329845
Ver en: http://hdl.handle.net/10261/329845
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oai:digital.csic.es:10261/329845

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329846
Dataset. 2022

ADDITIONAL FILE 2 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 2: Figure S2. a, Flow cytometry analysis of GFP populations in 23 wk-old Rosa26-RRAS2fl/flxSox2-Cre mouse spleen. Representative two-color contour plots of GFPhigh and GFPlow populations in total B cells (CD19+), CD5+ leukemic and CD23+ follicular B cells. Bottom, representation of GFP populations in T lymphocytes (CD3+). b, Percentage of GFPhigh cells in the indicated populations determined by flow cytometry. Data show means ± SEM from n = 8 mice (23 wk-old mice). ****p < 0.0001 (one-way ANOVA test). c, Western blot analysis of R-RAS2 expression of sorted GFPlow and GFPhigh leukemic cells from the spleen of a 25 wk-old Rosa26-RRAS2fl/flxSox2-Cre mouse (β-actin as loading control). d, Dot plot representation of GFPlow CD5+ leukemic B cell evolution in mb1-Cre mice over time, showing each mouse individually (n = 14). Data points were adjusted to a linear fit. These data were retrieved from the same mice as in Fig. 2i. e, Percentage of CD5+ cells in the indicated populations comparing GFPhigh and GFPlow distribution. Data show means ± SEM from n = 4 30 wk-old mice. Two-way ANOVA test. f, Heatmap of RNAseq expression data showing the genes differentially regulated in wild-type, follicular B cells (n = 6, 12wk-old), leukemic CD19 + CD5+ B cells (n = 6, 54wk-old), CD19+ GFPhigh (n = 2, 54wk-old) and CD19+ GFPlow (n = 2, 54wk-old) populations. Only genes significantly different between GFPhigh GFPlow populations (p < 0.05) and with a difference of 2-fold or more were used. Gene expression is shown in normalized log2 fold change., Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329846
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329846
HANDLE: http://hdl.handle.net/10261/329846
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329846
PMID: http://hdl.handle.net/10261/329846
Digital.CSIC. Repositorio Institucional del CSIC
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Ver en: http://hdl.handle.net/10261/329846
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329846

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329849
Dataset. 2022

ADDITIONAL FILE 3 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 3: Figure S3. a, Representative two-color contour plots of B cell populations in a peritoneal wash and the spleen of 12 wk-old mice according to the expression of the CD11b and CD5 markers in the CD19+ population. The blue square indicates CD11b + CD5- B1b cells in the peritoneum. Red square, the presence of CD11b + CD5+ B1a cells in control mice and leukemic cells. Quantification of CD11b + CD5+ cells is shown to the right in box and whiskers plots showing all points and median value. **p < 0.01; *** p < 0.001, two-tailed unpaired t-test with Welch’s correction. b, Representative two-color contour plots of IgM and GFP expression within the CD11b + CD5+ populations shown in a. Quantification of IgMbright cells within the CD11b + CD5+ B cell population is shown to the right in box and whiskers plots showing all points and median value. **** p < 0.0001, two-tailed unpaired t-test with Welch’s correction., Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329849
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329849
HANDLE: http://hdl.handle.net/10261/329849
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329849
PMID: http://hdl.handle.net/10261/329849
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329849
Ver en: http://hdl.handle.net/10261/329849
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oai:digital.csic.es:10261/329849

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329850
Dataset. 2022

ADDITIONAL FILE 4 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 4: Figure S4. a, Representative two-color contour plots of lymphoid populations in liver and spleen from 2 wk-old mice according to the expression of CD19 and CD5 and within the CD19 + CD5+ population according to the expression of CD21, B220, CD24, CD23 and CD38 markers. b, Column plots show the quantification of the percentage of CD19 + CD5+ B cells in liver and spleen bearing the markers shown in a. n = 4 mice per group. ** p < 0.01 ****p < 0.0001, ns, not significant (one-way ANOVA test)., Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329850
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329850
HANDLE: http://hdl.handle.net/10261/329850
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329850
PMID: http://hdl.handle.net/10261/329850
Digital.CSIC. Repositorio Institucional del CSIC
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Ver en: http://hdl.handle.net/10261/329850
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oai:digital.csic.es:10261/329850

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329851
Dataset. 2022

ADDITIONAL FILE 5 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 5: Figure S5. a, Principal component analysis of CD19 + CD21-CD23+ follicular B cells from Rosa26-RRAS2xmb1-Cre mice, CD19 + CD21-CD23+ follicular B cells from WT C57BL/6 J mice and of leukemic CD19 + CD5+, GFPlow and GFPhigh cells from Rosa26-RRAS2xmb1-Cre mice. b, Ingenuity Pathway Analysis (IPA) of differentially expressed genes associated with molecular mechanisms of cancer in leukemic versus normal follicular B cells. Pink-filled symbols: upregulated genes. Green-filled: downregulated genes. Double circle: protein complex; horizontal ellipse: transcription regulator; vertical ellipse: transmembrane receptor, diamond: enzyme; trapezium: transporter; triangle: phosphatase; inverted triangle: kinase; vertical rectangle: G protein-coupled receptor; circle: other. Black arrows: direct interactions; grey/white arrows: indirect interactions. Relationship labels: A: activation; B: binding; C: causation; CO: correlation; E: expression; EC: enzyme catalysis; I: inhibition; L: molecular cleavage; LO: localization; M: biochemical modification; miT: microRNA Targeting; P: phosphorylation/dephosphorylation; PD: protein-DNA binding; PP: protein-protein binding; PR: protein-RNA binding, RB: regulation of binding; RE: reaction; T: transcription; TR: translocation; UB: ubiquitination., Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329851
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329851
HANDLE: http://hdl.handle.net/10261/329851
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329851
PMID: http://hdl.handle.net/10261/329851
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329851
Ver en: http://hdl.handle.net/10261/329851
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329851

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329852
Dataset. 2023

ADDITIONAL FILE 5 OF IRON INDUCES RESISTANCE AGAINST THE RICE BLAST FUNGUS MAGNAPORTHE ORYZAE THROUGH POTENTIATION OF IMMUNE RESPONSES

  • Sánchez-Sanuy, Ferran
  • Mateluna-Cuadra, Roberto
  • Tomita, Keisuke
  • Okada, Kazunori
  • Sacchi, Gian Attilio
  • Campo, Sonia
  • San Segundo, Blanca
1 table., Additional file 5: Table S4. Expression data of defense-related genes in leaves of Control and High-Fe plants during M. oryzae infection (48 hpi)., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329852
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329852
HANDLE: http://hdl.handle.net/10261/329852
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329852
PMID: http://hdl.handle.net/10261/329852
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329852
Ver en: http://hdl.handle.net/10261/329852
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329852

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329853
Dataset. 2023

ADDITIONAL FILE 6 OF IRON INDUCES RESISTANCE AGAINST THE RICE BLAST FUNGUS MAGNAPORTHE ORYZAE THROUGH POTENTIATION OF IMMUNE RESPONSES

  • Sánchez-Sanuy, Ferran
  • Mateluna-Cuadra, Roberto
  • Tomita, Keisuke
  • Okada, Kazunori
  • Sacchi, Gian Attilio
  • Campo, Sonia
  • San Segundo, Blanca
1 table., Additional file 6: Table S5. Expression data (FPKM and Log2FC) of genes involved in the biosynthesis of diterpene phytoalexins and sakuranetin in leaves of Control and High-Fe plants (M. oryzae-infected and mock-inoculated plants)., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329853
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329853
HANDLE: http://hdl.handle.net/10261/329853
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329853
PMID: http://hdl.handle.net/10261/329853
Digital.CSIC. Repositorio Institucional del CSIC
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Ver en: http://hdl.handle.net/10261/329853
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329853

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329854
Dataset. 2022

ADDITIONAL FILE 6 OF OVEREXPRESSION OF WILD TYPE RRAS2, WITHOUT ONCOGENIC MUTATIONS, DRIVES CHRONIC LYMPHOCYTIC LEUKEMIA [DATASET]

  • Hortal, Alejandro
  • Oeste, Clara L.
  • Cifuentes, Claudia
  • Alcoceba, Miguel
  • Fernández-Pisonero, Isabel
  • Clavaín, Laura
  • Tercero, Rut
  • Mendoza, Pilar
  • Domínguez, Verónica
  • García-Flores, Marta
  • Pintado, Belén
  • Abia, David
  • García-Macías, Carmen
  • Navarro-Bailón, Almudena
  • Bustelo, Xosé R.
  • González, Marcos
  • Alarcón, Balbino
Additional file 6: Figure S6. a, Mutations found in human cancer involving the RRAS2 gene. Data obtained from cBioPortal (97,250 patients/100669 samples). Refseq: NM_012250. Ensembl: ENST00000256196. CCDS: CCDS7814. Uniprot: RRAS2_HUMAN. Missense mutations (green dots): 36. Truncating mutations (black dots): 6. Splice mutations (orange dots): 5. b, Quantification by RT-qPCR or total mouse (Rras2) and human (RRAS2) mRNA expression in purified splenic CD19+ B cells from Rras2(Q72L)fl/fl xmb1-Cre (Q72L) mice compared to purified B CD19+ B cells from control WT C57BL/6 mice and to CD19 + CD5+ leukemic B cells from Rosa26-RRAS2fl/flxmb1-Cre mice. Results show data obtained in triplicate normalized to the C57BL/6 control for n = 3 mice per group. All mice were 14 month-old. Data show means ± SEM for three mice per group. *p < 0.05; ns. Not significant (one-way ANOVA test). c, Left, quantification by flow cytometry of total B-cell number in spleens of 14 month-old control and Rras2(Q72L)fl/fl xmb1-Cre mice. Right, two-parameter flow cytometry plot showing frequency of IgM + CD5+ cells within CD19+ splenic B cells of control and Rras2(Q72L)fl/fl xmb1-Cre mice. d, Left, concentration of B-cells per microliter in blood of control and Rras2(Q72L)fl/fl xmb1-Cre mice. Right, two-parameter flow cytometry plot showing frequency of CD19 + CD5+ cells within blood B cells of control and Rras2(Q72L)fl/fl xmb1-Cre mice. e, Frequency of marginal zone (MZ) phenotype (CD21high, CD23low), and follicular (CD21low, CD23high) B cells within CD19+ splenic B cells of control and Rras2(Q72L)fl/fl xmb1-Cre mice. f, Phosflow cytometry analysis of different elements from PI3K-Akt-mTOR, Raf-Erk and proximal BCR signaling pathways. Wild-type CD19+ follicular B cells, CD19 + CD5+ leukemic cells from spleens of Rosa26-RRAS2fl/flxmb1-Cre mice and CD19+ non-leukemic B cells from Rras2(Q72L)fl/fl xmb1-Cre are shown. In grey, background fluorescence of the secondary antibodies. All mice were 23 wk-old. Data show means ± SEM from three mice per group. *p < 0.05; **p < 0.01; ****p < 0.0001 (one-way ANOVA test). g, Phosflow cytometry analysis of different elements from PI3K-Akt-mTOR, Raf-Erk and proximal BCR signaling pathways. CD19 + CD5+ leukemic cells from 30 wk-old Rosa26-RRAS2fl/flxmb1-Cre mice are compared with WT Control follicular (CD23highCD21−), marginal zone (MZ, CD23−CD21high), B1a (CD11b + CD5+) and B1b (CD11b + CD5−) spleen B cell populations. Data show means ± SEM from n = 3 mice per group. **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant (one-way ANOVA test)., Fundación Científica Asociación Española Contra el Cáncer Ministerio de Ciencia, Innovación y Universidades H2020 European Research Council Instituto de Salud Carlos III Consejería de Educación, Junta de Castilla y León, Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329854
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329854
HANDLE: http://hdl.handle.net/10261/329854
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329854
PMID: http://hdl.handle.net/10261/329854
Digital.CSIC. Repositorio Institucional del CSIC
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Ver en: http://hdl.handle.net/10261/329854
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oai:digital.csic.es:10261/329854

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329855
Dataset. 2023

ADDITIONAL FILE 7 OF IRON INDUCES RESISTANCE AGAINST THE RICE BLAST FUNGUS MAGNAPORTHE ORYZAE THROUGH POTENTIATION OF IMMUNE RESPONSES

  • Sánchez-Sanuy, Ferran
  • Mateluna-Cuadra, Roberto
  • Tomita, Keisuke
  • Okada, Kazunori
  • Sacchi, Gian Attilio
  • Campo, Sonia
  • San Segundo, Blanca
1 table., Additional file 7: Table S6. Expression data (FPKM and Log2FC) of genes involved in Fe homeostasis in leaves of Control and High-Fe plants (M. oryzae-infected and mock-inoculated plants)., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329855
Digital.CSIC. Repositorio Institucional del CSIC
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HANDLE: http://hdl.handle.net/10261/329855
Digital.CSIC. Repositorio Institucional del CSIC
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PMID: http://hdl.handle.net/10261/329855
Digital.CSIC. Repositorio Institucional del CSIC
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Ver en: http://hdl.handle.net/10261/329855
Digital.CSIC. Repositorio Institucional del CSIC
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Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329857
Dataset. 2023

ADDITIONAL FILE 8 OF IRON INDUCES RESISTANCE AGAINST THE RICE BLAST FUNGUS MAGNAPORTHE ORYZAE THROUGH POTENTIATION OF IMMUNE RESPONSES

  • Sánchez-Sanuy, Ferran
  • Mateluna-Cuadra, Roberto
  • Tomita, Keisuke
  • Okada, Kazunori
  • Sacchi, Gian Attilio
  • Campo, Sonia
  • San Segundo, Blanca
1 table., Additional file 8: Table S7. Statistics for RNAseq analysis of leaves from Control and High-Fe plants, mock-inoculated and M. oryzae-inoculated (48 hpi)., Consejo Superior de Investigaciones Cientificas (CSIC), Peer reviewed

Proyecto: //
DOI: http://hdl.handle.net/10261/329857
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329857
HANDLE: http://hdl.handle.net/10261/329857
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/329857
PMID: http://hdl.handle.net/10261/329857
Digital.CSIC. Repositorio Institucional del CSIC
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