Journal: Frontiers in Immunology
Article Title: From multi-omics to functional validation: the PTMRS stratifies TME and positions PDGFRB in CRC biology
doi: 10.3389/fimmu.2025.1728291
Figure Lengend Snippet: Impact of PDGFRb perturbation within tumor-associated fibroblasts on CRC cell phenotypes. (A) Western blots of t-PDGFRb and p-PDGFRb in CCD-18Co fibroblasts treated with vehicle (Veh), PDGF-BB, or PDGF-BB plus SUN. b-actin serves as a loading control. (B) Quantification of the p-PDGFRb/t-PDGFRb ratio relative to the Veh group. (C) Representative EdU staining of HCT116 cells cultured with CM from the three fibroblast conditions (Veh-CM, PDGF-BB-CM, SUN-CM). EdU (green), DAPI (blue), merged images as indicated. (D) Relative EdU labeling index in HCT116 cells. (E, F) CCK-8 assays showing relative cell viability of HCT116 (E) and DLD-1 (F) cells after 48 h exposure to Veh-CM, PDGF-BB-CM, or SUN-CM. (G) Representative Transwell images of migrated HCT116 and DLD-1 cells under the indicated CM conditions. (H, I) Quantification of migrated HCT116 (H) and DLD-1 (I) cells per 10× field. (J) Violin plots showing concentrations of CCL5, CXCL12, IL-6, IL-10, and TGF-b1 in fibroblast CM measured by ELISA. *P < 0.05, **P < 0.01, ***P < 0.001 vs Veh group; ns, no significant difference. Abbreviations: Veh, vehicle; SUN, sunitinib; CM, conditioned medium; PDGF-BB, the BB homodimer of platelet-derived growth factor.
Article Snippet: Human CRC cell line HCT116 and colonic fibroblast cell line (CCD-18Co) were obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37 °C in a 5% CO 2 humidified incubator.
Techniques: Western Blot, Control, Staining, Cell Culture, Labeling, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay