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. 2021 Feb 4;13(4):618. doi: 10.3390/cancers13040618

Figure 5.

Figure 5

Cyclin D1 expression was regulated by miR-224-5p. (A) Schematic of potential binding sites of miR-224-5p in the 3′UTR region of CCND1 mRNA. (B,C) GFP fluorescence in HEK293 cells after 48 h of co-transfection with indicated miRNA mimics (NC or miR-224-5p) and pEGFP-C1-CCND1-3′UTR wildtype/mutant plasmids. Scale bar, 100 nm. Fluorescent value of GFP (relative fluorescence) was normalized to concentration of total protein and shown as the fold of NC group. Data are mean ± s.e.m. ** p < 0.01; ns, no significant difference. (D) The regulation of cyclin D1 protein in RCC cells by miR-224-5p was determined by Western blot. (E) The regulation of CCND1 mRNA in RCC cells by miR-224-5p was determined by RT-qPCR. Data are mean ± s.e.m. * p < 0.05; ** p < 0.01; *** p < 0.001. (Original Western blots see Figure S7).