Fig. 2

miR-208a affected the levels of the p21/AKT/mTOR pathway in A549 cells. A549 cells were transfected with the miR-208a mimics, negative control miRNA (miRNA-NC), miR-208a inhibitor or inhibitor-NC for 24 h before lysis. a Western blot analysis the expression levels of p21. b Putative miR-208a binding sequences in the 3′-UTR of p21 mRNA. c Full-length 3′-UTR of p21 and the mutant sequences were cloned into the downstream of the luciferase reporter gene named as pGL3-UTR-WT and pGL3-UTR-MUT, respectively. Each of the plasmids and pRL-TK, were cotransfected into A549 cells, either with miR-208a mimics or inhibitor as indicated. Luciferase activity was assayed 48 h after transfection. All experiments were repeated three times independently. The results are presented as the means ± SEM (n = 3). *P < 0.05. d The membranes were incubated with antibodies to AKT, p-AKT, mTOR, or p-mTOR. The samples were derived from the same experiment and analyzed under the same experimental conditions