Fig. 4

PDK2 is a direct target of miR-149-3p. a Schematic diagram of the protocol used to search for candidate target genes which are predictively regulated by miR-149-3p. b HCT-8 and HCT116 cells were transiently transfected with an miR-149-3p mimic or inhibitor. The mRNA levels of PDK2 and HK1 were analyzed by quantitative real-time PCR. c Diagram of the miR-149-3p putative binding sites in the 3′-UTR of PDK2. The mutant sequences used in the luciferase reporters are indicated in red. d Human embryonic kidney 293 T cells and HCT116 cells were cotransfected with luciferase reporter constructs and the miR-149-3p mimic (50 nmol/L) or miRNA control for 48 h. Relative luciferase activity data were normalized to corresponding control. e HCT-8 and HCT116 cells were transfected with either the miR-149-3p mimic or inhibitor for 48 h, and the expression of PDK2 was determined by Western blot analysis. f HCT116 cells transfected with the miR-149-3p inhibitor or anti-NC were treated with or without DCA for 24 h. The expression of PDK2 was determined by Western blot analysis. g The expression of PDK2 and p-PDHA1 in chemosensitive and chemoresistant CRC cells were analyzed by Western analysis. Data shown are representative pictures of three experiments. The results of three independent experiments performed in triplicate are shown as the mean ± SEM. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001