Figure 5

MiR875-5p inhibits Pten and regulates phosphorylation of Akt. (a) Schematic of reporter construct used to evaluate the role of complementarity between miR875-5p and Pten 3′ UTR (blue). The wild-type (WT) construct had a 22-nt fragment of the Pten UTR, which included miR875-5p target sites (gray) inserted within the firefly luciferase 3′ UTR. The mutant construct was identical to the WT construct, except that it had several disruptions of pairing to miR875-5p. (b) qRT-PCR analysis of Pten in mDP cells cultured with the control or mimic miR875-5p after transfection and normalization to Gapdh expression. Western blotting results of PTEN and GAPDH in mDP cells cultured with the control or transfected with mimic miR875-5p. The ful-length blots are presented in Supplementaly Fig. 1a. (c) Pten WT and mutation reporter activities were evaluated in mDP cells transfected with mimic miR875-5p. (d) Western blotting of p-Akt and Akt in mDP cells with or without mimic miR875-5p after culturing for 1 hour. The ful-length blots are presented in Supplementaly Fig. 1b.*P < 0.05. Error bars represent the mean ± S.D.