Fig. 4

Targeting miR-4477b inhibited the proliferation, clone formation, and induced apoptosis in renal carcinoma cells. A The relative expression of miR-4477b and miR-514b was tested in tumor and normal kidney cell lines. N = 5. B The relative expression of miR-4477 was tested in tumor tissue and adjacent normal tissue in patient samples. N = 10. C The levels of cellular glucose (left) and lactate (right) in Caki-1 and 769-P cells. N = 3. D Colony formation assay was performed in Caki-1 and 769-P cells, transfected with miR-4477b antisense inhibitor or scramble inhibitor. D After transfected with miR-4477b antisense inhibitor or scramble inhibitor, the cell viability of Caki-1 and 769-P cells was tested at indicated time points by CCK8 assay. E The level of apoptosis of Caki-1 and 769-P cells transfected with miR-4477b antisense inhibitor or scramble inhibitor was analyzed by Annexin V and PI staining by flow cytometry. F The relative expression of indicated genes was test by QPCR in Caki-1 cells transfected with miR-4477b antisense inhibitor or scramble inhibitor. G Upper panel: the WT and mutation of the 3′-UTR region of TEF gene. The mutation of the binding site of miR-4477b was shown in red. Lower panel: Hek293 cells were transfected with the WT and MUT TEF 3′-UTR-Luc vector together with Renilla luciferase and miR-4477b mimic. Cells were harvested for luciferase assays 48 h after transfection. The relative luciferase activity was normalized by Renilla luciferase activity. *P < 0.05