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: CSF1R inhibition promotes CAF activation and extracellular matrix remodeling (A) GO enrichment analysis of upregulated genes in PLX3397-treated tumors. (B) Heatmap of tumor stiffness measured by atomic force microscopy (AFM). (C) Quantification of tumor stiffness (kPa) in control and PLX3397-treated groups ( n = 5 per group). (D and E) Representative Masson’s trichrome, picrosirius red, and IHC staining for collagen I and collagen III in xenograft tumors treated with PLX3397: (D) CT26 and (E) CMT93. Scale bar, 60 μm. (F) Experimental design for in vivo evaluation of PLX3397 combined with EDHB in BALB/c mice injected with CT26 cells. (G–I) Tumor growth in control and EDHB-treated CT26 xenografts with or without PLX3397 ( n = 5 per group): (G) growth curves, (H) tumor weight ( n = 5 per group), and (I) representative tumor images. (J) Representative collagen staining (picrosirius red, Masson’s trichrome, and Collagen III) in control and EDHB-treated groups. Scale bar, 60 μm. (K) Quantification of (J). (L) Representative IHC of CD8+T cell in control and EDHB-treated groups ( n = 5 per group). Scale bar, 60 μm. (M–O) Tumor growth in xenografts derived from co-inoculation of CT26 cells with NIH/3T3 fibroblasts, treated with or without PLX3397 ( n = 5 per group); (M) tumor images, (N) growth curves, and (O) tumor weight. Unless specified otherwise, data were presented as the means ± SEM. t tests were used in (C), (K), and (L). One-way ANOVA tests were used in (G), (H), (N), and (O) to determine statistical significance. ∗ 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: Inhibition, Activation Assay, Microscopy, Control, Immunohistochemistry, In Vivo, Injection, Staining, Derivative Assay

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

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: Targeting CTGF enhances the anti-tumor efficacy of CSF1R inhibition in CRC (A) Western blot analysis of Ctgf levels in normal control (NC) and Ctgf-KO CT26 cells. (B–D) CT26 control or Ctgf-KO cells were subcutaneously implanted into BALB/c mice and treated with PLX3397 ( n = 6). (B) Tumor growth curves, (C) representative tumor images, and (D) tumor weights ( n = 6). (E and F) Representative images (E) and quantification (F) of Masson’s trichrome, picrosirius Red, and Collagen III staining in CT26 tumors from control and Ctgf-KO groups ( n = 6). Scale bar, 60 μm. (G) Representative IHC images (left) and quantification (right) of α-SMA-positive fibroblasts in CT26 tumor sections ( n = 6). Scale bar, 60 μm. (H) IHC images (left) and quantification (right) at tumor margin and core ( n = 6). Scale bar, 60 μm. (I) Representative flow cytometry plots (left) and quantification (right) of CD8 + T cells ( n = 6). (J–M) CT26 cells were implanted and treated with PLX3397, anti-CTGF, or the combination ( n = 6). (J) Experimental design, (K) tumor growth curves, (L) representative images, and (M) tumor weights ( n = 6). (N and O) Representative images (left) and quantification (right) of Masson’s trichrome, picrosirius Red, Collagen III (N), and α-SMA IHC (O) in tumors from control and anti-CTGF-treated groups ( n = 6). Scale bar, 60 μm. (P and Q) Assessment of CD8 + T cell infiltration in anti-CTGF-treated tumors. (P) IHC images (left) and quantification (right) at tumor margin and core ( n = 6). Scale bar, 60 μm. (Q) Representative flow cytometry plots (left) and quantification (right) ( n = 6). Unless specified otherwise, data were presented as the means ± SEM. t tests were used in (F), (G), (H), (I), (N), (O), (P), and (Q). One-way ANOVA tests were used in (B), (D), (K), and (M) to determine statistical significance. ∗ 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: Inhibition, Western Blot, Control, Staining, Flow Cytometry