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. 2021 May 3;12(12):3726–3740. doi: 10.7150/jca.48418

Figure 5.

Figure 5

miR-146b-3p negatively regulates MAP3K10 by binding to the MAP3K10 3'-UTR. A) Sequence alignment of miR-146b-3p with the 3'-UTR of MAP3K10. The seed sequence of let-7a (top) matches the 3'-UTR of MAP3K10. Bottom, mutations of the 3'-UTR of MAP3K10 for creating the Renilla reporter constructs (the mutant nucleotides of the miR-146b-3p binding site are red and italic).B), C), and D), The comparison of MAP3K10 expression between matched normal pancreatic tissue and pancreatic cancer tissues in 12 patients. (B) Analysis of the gene expression levels of MAP3K10 by real time PCR and (right) the data were analyzed using the Mann-Whitney U test. (C) Analysis of the protein expression levels of MAP3K10 by western blotting and (D) the data were analyzed using the Mann-Whitney U test. T, pancreatic cancer tumor tissue; N, pair-matched adjacent non-tumor tissue. E) Correlation of the protein expression of MAP3K10 with miR-146b-3p expression in 12 pairs of pancreatic cancer tissues and matched normal pancreatic tissues. Coefficient of determination from logarithmic regression model (R2) and P values are given (R2=0.7004; P<0.0001). F) Analysis of the protein expression levels of MAP3K10 in six pancreatic cancer cell lines by western blotting. G) Analysis of Gene expression of MAP3K10 in Mia PaCa-2 and BxPC-3 cells after transfection with miR-146b-3p mimic or mimic NC or no transfection by real time PCR.H) Analysis of protein expression of MAP3K10 in Mia PaCa-2 and BxPC-3 cells after transfection with miR-146b-3p mimic or mimic NC or no transfection by western blot. I) Analysis of luciferase activity. miR-146b-3p mimic inhibited wild type but not mutant MAP3K10 3'-UTR reporter activity. *P <0.05, compared to the mimic NC group. P values were obtained by two-tailed Student's t test.