Journal: Cell Reports Medicine
Article Title: Targeting CTGF overcomes resistance to CSF1R inhibitors by preventing CAF activation in colorectal cancer
doi: 10.1016/j.xcrm.2025.102480
Figure Lengend Snippet: CRC cell-derived CTGF promotes CAF activation and ECM remodeling in vivo and in vitro (A) Representative immunoblots of Ctgf in the control and Ctgf overexpression CT26 and CMT93 cells. (B and C) Schematic diagram for establishing syngeneic tumor-bearing mouse model in BALB/c mice with indicated CT26 cells ( n = 5 per group). Tumor growth curve (left panel) and representative images of tumors (right panel). (C) Tumor weight for indicated groups as presented in (B) ( n = 5). (D) Representative images (left panel) and quantification (right panel) of Masson’s, picrosirius red, Collagen III, and α-SMA staining of tumor tissues for control and Ctgf overexpression groups from CT26 cell-derived syngeneic mouse model ( n = 5). Scale bar, 60 μm. (E) Representative images of CD8 + T cell staining at CT26 tumor margin and in the tumor core (bottom panels). Representative images (left panel) and quantification (right panel) ( n = 5). Scale bar, 100 μm. (F and H) Representative immunofluorescence images for α-SMA and Collagen I in NIH/3T3 treated with supernatants of CT26 or dosage rectification. Scale bar, 100 μm. (G and I) NIH/3T3 cells were treated with a gradient dose of rCTGF or supernatants from Ctgf overexpression CT26 and CMT93 cells. The α-SMA and collagen I expression levels were detected by western blotting analysis. (J) The ability of migration and invasion was performed by Transwell assays ( n = 3 biological replicates); scale bar, 100 μm. (K and L) CT26 cells or Ctgf overexpression cells were injected subcutaneously into BALB/C alone or mixed with NIH/3T3 cells in a 1:2 ratio ( n = 5). Tumor growth curves (K), tumor weight (left panel) ( n = 5), and tumor image (right panel) (L) in each treatment group. (M) Representative IHC staining of α-SMA-positive fibroblasts (top row) and quantification of α-SMA-positive fibroblasts (right panel) in (K) ( n = 5). Scale bar, 60 μm. Immunofluorescent staining and quantification of CD8 + T cells (bottom row) in (K); scale bar, 100 μm. Unless specified otherwise, data were presented as the means ± SEM. t tests were used in (B), (C), (D), (E), (J), (K), (L), and (M). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗ p < 0.0001; ns, not significant.
Article Snippet: Anti-Collagen Type III Rabbit mAb , Proteintech , Cat# 22734-1-AP; RRID: AB_2879158.
Techniques: Derivative Assay, Activation Assay, In Vivo, In Vitro, Western Blot, Control, Over Expression, Staining, Immunofluorescence, Expressing, Migration, Injection, Immunohistochemistry