Fig. 2

NF-κB activation could negatively regulate the expression of miR-195-5p/497–5p in CCSCs. a, Prediction of binding sites in the miR-195-5p and miR-497-5p promoter region. b, Dual-luciferase reporter assay results showing the relative luciferase activity after deletion mutation. c, The protein expression of p65 and extent of p65 phosphorylation detected by Western blot analysis after treatment with overexpressed or downregulated p65 in CCSCs. d, The activity of NF-κB and that of miR-195-5p/497–5p promoter after CCSCs were treated with p65 detected by luciferase assay. e, The expression of miR-497-5p and miR-195-5p after CCSCs were treated with p65 detected by RT-qPCR. f, The expression of miR-497-5p and miR-195-5p in nude mice tumors was detected by RT-qPCR. g, The protein expression of p65 and extent of p65 phosphorylation in tumor tissues in response to different treatments after 24 h detected by Western blot analysis. Measurement data were expressed as mean ± s.d. Data between two groups were compared using unpaired t-test. Data among multiple groups were compared using one-way analysis of variance and then analyzed with Tukey’s post-hoc test. Six mice in each group. Cell experiment was repeated three times. * p < 0.05 vs. control