Fig. 4

TLR4 as a target gene of miR-5195-3p in CRC cells. (A) Schematic of miR-5195-3p putative binding sites in WT 3ʹ UTR of TLR4 and MUT 3′-UTR of TLR4. (B-C) Relative luciferase activity of TLR4 3′-UTR in SW1116 and HT29 cells co-transfected with the indicated reporters and miR-5195-3p mimics oligonucleotides or miR-NC. (D) Quantitative real time PCR analysis of the mRNA levels of TLR4 in SW1116 and HT29 cells transfected with miR-5195-3p mimics or miR-NC. (E) Western blot analysis of the protein levels of TLR4 in SW1116 and HT29 cells transfected with miR-5195-3p mimics or miR-NC. (F) The mRNA level of TLR4 was detected by quantitative real time PCR in tumor and adjacent tissues derived from CRC patients. (G) Spearman’s analysis of the correlation between miR-5195-3p and mRNA levels of TLR4 mRNA expression in human CRC tissues. Data were expressed as means ± SD of three independent experiments. **p < 0.01, ***p < 0.001, compared with miR-NC