Journal: Cell Death & Disease
Article Title: FGFR4 promotes CAF activation through the CXCL10-CXCR3 axis in colon cancer
doi: 10.1038/s41419-025-07588-y
Figure Lengend Snippet: A Collagen gel contraction assay results show the effect of recombinant CXCL10 protein on the contractility of NIH/3T3 cells (n = 6/group). Representative gel images are shown together with the quantification of collagen gel contraction. B Migration and invasion assay results show the effect of recombinant CXCL10 protein on the migration and invasion properties of NIH/3T3 cells. The effect of CXCL10 was compared to that of TGFβ, which induces CAF activation. C The effect of recombinant CXCL10 protein (ng) on CAF marker gene levels in NIH/3T3 cells was determined using western blotting and RT-qPCR analyses. D Effect of Cxcl10 knockdown on CAF marker gene expression. NIH/3T3 cells were incubated for 24 h with conditioned media (CM) obtained from siNC- or siCXCL10-transfected CT-26/FGFR4 cells. Western blotting and RT-qPCR analyses of CAF marker expression in NIH/3T3 cells. E Effect of CXCL10 neutralizing antibody on CAF marker gene expression. NIH/3T3 cells were incubated in CM obtained from CT-26/FGFR4 cells in the presence of either normal anti-IgG or anti-CXCL10 neutralizing antibodies (concentrations shown in μg/mL) for 24 h. F Effect of AMG487, a pharmacological CXCR3 inhibitor, on gene expression of CAF markers. The CM of CT-26/FGFR4 cells, alone or in combination with the CXCR3 inhibitor, was used to treat the NIH/3T3 cells for 24 h. G Invasion assay results depicting the effect of the CXCR3 inhibitor AMG487 (2 μM) on the invasion capabilities of CT-26 primary tumor-derived CAFs. H Invasion assay results show the effects of various CXCR3 inhibitors on the invasive properties of CT26/FGFR4 tumor-derived CAFs. The CXCR3 inhibitors used (at 2 μM) were: TAK779, NBI74330, and SCH546738. Data are presented as mean ± SD. Statistical significance: *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 using Student’s t -test and one-way ANOVA). NT no treatment, AMG AMG487, TAK TAK779, NBI NBI74330, SCH SCH54673 .
Article Snippet: The following inhibitors were used: BLU9931 (Merck, Kenilworth, NJ, USA; 538776), AMG487 (MedChemExpress, Monmouth Junction, NJ, USA; HY-15319), TAK779 (MedChemExpress; HY-13406), NBI74330 (MedChemExpress; HY-15320), and SCH546738 (MedChemExpress; HY-10017).
Techniques: Collagen Gel Contraction Assay, Recombinant, Migration, Invasion Assay, Activation Assay, Marker, Western Blot, Quantitative RT-PCR, Knockdown, Gene Expression, Incubation, Transfection, Expressing, Derivative Assay