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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Exosome-mediated transfer of miR-93-5p from cancer-associated fibroblasts confer radioresistance in colorectal cancer cells by downregulating FOXA1 and upregulating TGFB3

Fig. 6

miR-93-5p promotes the activation of TGF-β signaling pathway by inhibiting FOXA1. SW480 cells were transfected with FOXA1 overexpression plasmid and co-cultured with miR-93-5-exo. a TGFB3 mRNA expression in CRC tissues (n = 75) and adjacent normal tissues (n = 75) determined by RT-qPCR, normalized to GAPDH. b TGFB3 protein band pattern in CRC tissues (n = 75) and adjacent normal tissues (n = 75) detected by Western blot analysis. c TGFB3 protein expression in CRC tissues (n = 75) and adjacent normal tissues (n = 75) determined by Western blot analysis, normalized to GAPDH. d Binding of FOXA1 to TGFB3 promoter detected by dual luciferase reporter gene assay. e–f Enrichment of FOXA1 in the promoter region of TGFB3 detected by ChIP assay. g Analysis of the correlation between FOXA1 and TGFB3 expression in CRC. h Detection of nuclear translocation of TGFB3 by immunofluorescence staining (400 ×). Green represented TGFB3 protein, red referred to cells and yellow referred to expression of TGFB3 in the nucleus. i mRNA expression of FOXA1, TGFB3, TGF-β1, and Smad3 in SW480 cells determined by RT-qPCR, normalized to GAPDH. j–k Protein bands and protein levels of FOXA1, TGFB3, TGF-β1, and Smad3 in SW480 cells detected by Western blot analysis, normalized to GAPDH. Values obtained from three independent experiments in triplicate are expressed as mean ± SD and analyzed by one-way ANOVA followed by Tukey’s post hoc test among multiple groups. *p < 0.05 compared with SW480 cells treated with oe-NC or NC-exo + oe-NC

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