Fig. 2: miR-4779 down-regulates PAK2 and CCND3 by direct targeting

a, b HCT116 cells were transfected with miR-NC or miR-4779 for 24 h. The mRNA levels of PAK2 and CCND3 were determined by qRT-PCR a. HPRT1 was used as a normalizer. Data show the relative each gene expression in miR-4779 transfected samples as compared with a miR-NC transfected samples. The expression of mature miR-4779 was determined by qRT-PCR b. U6 snRNA was used as a normalizer. c HCT116, A549, H460, MCF7, and HT-29 cells were transfected with miR-NC or miR-4779 for 48 h, and the protein levels of PAK2 and CCND3 were determined by western blotting using the corresponding antibodies. d-e Structure of reporter constructs containing 3’UTRs of PAK2 d and CCND3 e downstream of the luciferase ORF. The aligned sequences of constructs (red) complementary to the seed sequence of the miR-4779 (bold) and the mutant sequences are shown. The mutated regions are shaded. HCT116 cells were transfected with a combination of the indicated reporters and miRNAs, and luciferase assay was performed 48 h after transfection f. The fold change in relative luciferase activity in transfected cells with miR-NC was set as 1. Data are presented as averages of triplicate measurements with error bars representing standard deviations. ^**P < 0.01 and ^***P < 0.001