Fig. 2

MiR-34c inhibits SOX4 expression. a Evaluation of miR-34c targets: the Venn diagram was generated by combining miRWalk-predicted miR-34c targets and the upregulated NPC genes from Shi et al., 2006 [69] using the online tool at www.bioinformatics.psb.ugent.be/webtools/Venn. b and c qRT-PCR of genes highly expressed in C666–1 cells compared to NP69/NP460 cells. b C666–1 cells were transiently transfected with pre-miR-34c (20 or 50 nM) for 72 h. c C666–1 cells were treated with SB431542 (10 or 20 μM) for 72 h. d Relative luciferase activity after transient transfection with pre-miR-34c (20 nM) for 48 h, followed by co-transfection with Renilla plasmid (100 ng) and either pMIR-SOX4 3’UTR Wildtype (WT) (150 ng) or pMIR-SOX4 3’UTR Mutant (150 ng) for 24 h. e qRT-PCR for SOX4 in NP69 cells transiently transfected with miR-control (50 nM), or pre-miR-34c (20 or 50 nM) for 72 h; 8 h after transfection, the media was changed to MEM 0.5% FBS; the following morning, cells were treated with recombinant TGFβ1 (10 ng/mL) for 48 h. f Kaplan-Meier plot of DRFS for NPC patients (n = 53) dichotomized based on low (<median) vs. high (>median) SOX4 mRNA expression (median follow-up time = 6 years). The data are represented as the mean ± SEM of at least three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001