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. 2022 Apr 15;8:205. doi: 10.1038/s41420-022-00932-7

Fig. 2. miR-15a-5p targets CHEK1 and downregulates CHEK1 expression.

Fig. 2

A Schematic diagram of miR-15a-5p binding sites in the 3′UTR of CHEK1 mRNA along with mutation site. B The relative luciferase activity of CHEK1-3′UTR-reporters responding to the miR-15a-5p mimic. The reporters were co-transfected with miR-15a-5p mimic into HEK-293T. Luciferase activity was tested after transfected for 48 h. C miR-15a-5p expression in HuCCT1 and HuH28 cells determined by RT-qPCR. D Western blot assay of CHEK1 protein in HuCCT1 and HuH28 cells. Quantitative results of protein bands are displayed at the right of the panel. In panels C and D, Mimic-NC, miR-15a-5p mimic, inhibitor-NC or miR-15a-5p inhibitor were individually transfected into HuCCT1 and HuH28 cells, RNA and protein levels of cells were analyzed after transfection for 48 h. The data are presented as mean ± standard deviation. Comparison between two groups was conducted using unpaired t-test, and the values at multiple groups were compared using one-way ANOVA followed by Tukey’s post-hoc tests. *means comparison against the NC or mimic-NC group, P < 0.05; #means comparison against inhibitor-NC group, P < 0.05. The cell experiments were repeated three times independently.