Resultados de investigación

Supplementary Material 4: Fig. 4. A. Stain-free gels (top) were used to evaluate the binding of anti-TGFα antibodies to the cytotoxic agents DM1, MMAF, and DXd. A change in the migration of the antibody in the gel was observed after coupling. Bottom blots: one hundred nanograms of each ADC and the unconjugated mAbs were loaded in 12% SDS-PAGE gels. The cytotoxic payload bound to the light and heavy chains of the ADCs was analyzed by Western blot using anti-DM1, anti-MMAF, and anti-DXd antibodies, as shown below. B. Detection of DM1 bound to 5F1 or 16B10 mAbs, compared to T-DM1. Equal amounts of the nude or coupled antibodies were used. Blots were analyzed using anti-DM1 antibodies. C. Restoration of 16B10-MMAF sensitivity in a TGFA-KO clone. The parental cell line IMIM-PC2, the CRISPR/Cas9 clone #25 and the TGFA reconstituted clone #13 were seeded in 6-well plates and treated with 2.5 nM of the ADCs (colored boxes). After 5 days, the effect on cell proliferation was assessed by cell counting. The results obtained are represented as the mean ± SD of the triplicates of an experiment that were repeated three times. **P < 0.01, calculated by Mann-Whitney U test., Predoctoral contract Cancer Research Foundation of the Salamanca University (FICUS) Instituto de Salud Carlos III through a Miguel Servet program Ministry of Economy and Competitiveness of Spain Instituto de Salud Carlos III through CIBERONC Junta de Castilla y León ALMOM ACMUMA UCCTA CRIS Cancer Foundation and the Regional Development Funding Program (FEDER) “A way to make Europe”, Peer reviewed

Supplementary Material 5: Fig. 5. A. IC50 ± SD of the 16B10-MMAF ADC or free drug in each pancreatic cancer cell line, including CRISPR/Cas9 clones. All values were calculated using GraphPad Prism 8 software. B. Graphical representation of the values shown in A., Predoctoral contract Cancer Research Foundation of the Salamanca University (FICUS) Instituto de Salud Carlos III through a Miguel Servet program Ministry of Economy and Competitiveness of Spain Instituto de Salud Carlos III through CIBERONC Junta de Castilla y León ALMOM ACMUMA UCCTA CRIS Cancer Foundation and the Regional Development Funding Program (FEDER) “A way to make Europe”, Peer reviewed

This dataset compiles records of Macrobrachium nipponense occurrences across its non-native range, including geographical and bibliographical information. It supports the article “A new non-native species detected through citizen science: the presence of a Macrobrachium species (Decapoda, Palaemonidae) in the Guadalquivir River estuary (SW Spain), with an overview of its global spread.” The Excel file contains 16 fields: Record, Continent, Country, CountryCode, Location, Latitude, Longitude, Sampling year, Record date, Number of Individuals, Habitat, Substrate, Reference, gbifID, and occurrenceID. Records include detailed metadata on sampling site, estimated coordinates, year of detection, habitat type, and original sources. Some coordinates were inferred based on location descriptions and are marked as such. This dataset aims to facilitate future assessments of the biogeographical expansion of M. nipponense and inform monitoring and management strategies targeting non-native freshwater decapods. It may be used for comparative invasion biology studies, geospatial analyses, or integration into global biodiversity platforms., Agencia Estatal de Investigación: PTA2022-021378-I; Junta de Andalucía: DGP_POST_2024_00956, Peer reviewed

Supplementary Material 6: Fig. 6. Internalization of anti-TGFα-MMAF antibodies in pancreatic cancer cell lines. Capan-1 cells were treated for 12 hours at 37°C with 2.5 nM of 5F1-MMAF or 16B10-MMAF, with or without prior incubation with 50 µM of chloroquine (Ch), 3 hours before. White dots indicate colocalization between the anti-TGFα ADCs and LAMP1, which is represented by scatter graphs at the bottom. Scale: 25 µm. Anti-TGFα ADCs: red, LAMP1: green; DAPI: blue., Predoctoral contract Cancer Research Foundation of the Salamanca University (FICUS) Instituto de Salud Carlos III through a Miguel Servet program Ministry of Economy and Competitiveness of Spain Instituto de Salud Carlos III through CIBERONC Junta de Castilla y León ALMOM ACMUMA UCCTA CRIS Cancer Foundation and the Regional Development Funding Program (FEDER) “A way to make Europe”, Peer reviewed

Supplementary Material 7: Fig. 7. A. Expression levels of total and active EGFR pancreatic cancer cell lines. One milligram of total protein from each cell line was immunoprecipitated with an anti-EGFR antibody to assess both the total amount and the phosphorylation status of EGFR. Calnexin was used as a loading control. B. Relationship between TGFA and EGFR expression in pancreatic cancer. Data was obtained from the TNMplot online tool., Predoctoral contract Cancer Research Foundation of the Salamanca University (FICUS) Instituto de Salud Carlos III through a Miguel Servet program Ministry of Economy and Competitiveness of Spain Instituto de Salud Carlos III through CIBERONC Junta de Castilla y León ALMOM ACMUMA UCCTA CRIS Cancer Foundation and the Regional Development Funding Program (FEDER) “A way to make Europe”, Peer reviewed

Predoctoral contract Cancer Research Foundation of the Salamanca University (FICUS) Instituto de Salud Carlos III through a Miguel Servet program Ministry of Economy and Competitiveness of Spain Instituto de Salud Carlos III through CIBERONC Junta de Castilla y León ALMOM ACMUMA UCCTA CRIS Cancer Foundation and the Regional Development Funding Program (FEDER) “A way to make Europe”, Peer reviewed

Peer reviewed

Understanding the trophic ecology of herbivorous and detritivorous fishes is essential for evaluating their ecological roles in coral reef ecosystems. In this study, we combined bulk stable isotope (δ15N and δ13C) and fatty acid analyses to investigate trophic partitioning and dietary resource use among herbivorous and detritivorous fishes from the Great Barrier Reef, Australia. Isotopic niches and fatty acid profiles confirmed significant trophic partitioning among algivores, detritivorous surgeonfishes, and parrotfishes. We also applied mixing models based on these ecological tracers to quantify the contributions of basal dietary sources to the fish. Our findings further support previous dietary knowledge for several species, including algivorous acanthurids, kyphosid chubs, and the rabbitfish Siganus doliatus. However, they also highlight trophic niche specializations within these groups, particularly in Naso unicornis, which assimilates substantial dietary protein from epiphytic cyanobacteria despite a macroalgal diet, and in the detritivorous Ctenochaetus striatus, which exhibited isotopic similarities to parrotfishes but differed in fatty acid composition, likely due to a higher intake of diatoms. Additionally, our analyses reinforce the distinctive dietary composition of parrotfishes, emphasizing the complexity of their feeding biology, in which microscopic photoautotrophs such as cyanobacteria and dinoflagellates play a key dietary role that has often been overlooked in previous studies on their nutritional ecology. Furthermore, these findings underscore the usefulness of multi-tracer approaches in refining our understanding of coral reef fish trophic ecology., Peer reviewed

[References:] Byrne, Maria (1990): Annual reproductive cycles of the commercial sea urchin Paracentrotus lividus from an exposed intertidal and a sheltered subtidal habitat on the west coast of Ireland. Marine Biology, 104(2), 275-289, https://doi.org/10.1007/BF01313269 Lozano, J; Galera, J; López,, S; Turon, X; Palacin, C (1995): Biological cycles and recruitment of Paracentrotus lividus (Echinodermata: Echinoidea) in two contrasting habitats. Marine Ecology Progress Series, 122, 179-191, [Coverage:] Median Latitude: 27.980300 * Median Longitude: -15.656640 * South-bound Latitude: 27.760200 * West-bound Longitude: -15.829200 * North-bound Latitude: 28.150400 * East-bound Longitude: -15.414600, Biometric and reproductive data of three sea urchin common species, Paracentrotus lividus, Arbacia lixula, and Sphaerechinus granularis, in the Central-Eastern Atlantic's rocky shores were collected over a one-year sampling period, from June 2020 to May 2021, in different localities of Gran Canaria Island. The datasets include biometric measurements such as test diameter, height, total wet weight, gut weight, and gonad weight, along with reproductive data including sex and maturity stage for each individual sampled at the different localities. In addition, oocytes from mature females of each species were measured to compare the size between species. The collected specimens were identified in the laboratory to the lowest possible taxonomic level, and those selected for sex and sexual maturity analysis were examined following a methodology based on macroscopic observation of the gonads and histological analysis (Byrne, 1990; Lozano et al., 1995). Biometric measurements, such as test diameter, height, total wet weight, gut weight, and gonad weight, along with reproductive data, including sex and maturity stage for each individual sampled at the different localities, were collected. The collected specimens were identified in the laboratory to the lowest possible taxonomic level, and those selected for sex and sexual maturity analysis were examined following a methodology based on macroscopic observation of the gonads and histological analysis (Lozano et al., 1995)., Xunta de Galicia, grant/award no. IN606B-2024/018: Interacciones tróficas y procesos ecosistémicos en peces mesopelágicos (TROF-MP), Peer reviewed

[References:] Byrne, Maria (1990): Annual reproductive cycles of the commercial sea urchin Paracentrotus lividus from an exposed intertidal and a sheltered subtidal habitat on the west coast of Ireland. Marine Biology, 104(2), 275-289, https://doi.org/10.1007/BF01313269 Lozano, J; Galera, J; López,, S; Turon, X; Palacin, C (1995): Biological cycles and recruitment of Paracentrotus lividus (Echinodermata: Echinoidea) in two contrasting habitats. Marine Ecology Progress Series, 122, 179-191 Rueden, Curtis T; Schindelin, Johannes; Hiner, Mark C; DeZonia, Barry E; Walter, Alison E; Arena, Ellen T; Eliceiri, Kevin W (2017): ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics, 18(1), 529, https://doi.org/10.1186/s12859-017-1934-z, [Coverage:] Median Latitude: 27.980300 * Median Longitude: -15.656640 * South-bound Latitude: 27.760200 * West-bound Longitude: -15.829200 * North-bound Latitude: 28.150400 * East-bound Longitude: -15.414600, Biometric and reproductive data of three sea urchin common species, Paracentrotus lividus, Arbacia lixula, and Sphaerechinus granularis, in the Central-Eastern Atlantic's rocky shores were collected over a one-year sampling period, from June 2020 to May 2021, in different localities of Gran Canaria Island. The datasets include biometric measurements such as test diameter, height, total wet weight, gut weight, and gonad weight, along with reproductive data including sex and maturity stage for each individual sampled at the different localities. In addition, oocytes from mature females of each species were measured to compare the size between species. The collected specimens were identified in the laboratory to the lowest possible taxonomic level, and those selected for sex and sexual maturity analysis were examined following a methodology based on macroscopic observation of the gonads and histological analysis (Byrne, 1990; Lozano et al., 1995). After histological analysis, the ovaries of mature females of three sea urchin common species, Paracentrotus lividus, Arbacia lixula, and Sphaerechinus granularis, on the Central-Eastern Atlantic's rocky shores were photographed. The oocytes were measured using ImageJ software (Rueden 2017) to compare the sizes of these cells between species. The dataset includes the diameter of each oocyte measured and the code of each individual., Xunta de Galicia, grant/award no. IN606B-2024/018: Interacciones tróficas y procesos ecosistémicos en peces mesopelágicos (TROF-MP), Peer reviewed