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. 2012 Mar 19;109(14):5316–5321. doi: 10.1073/pnas.1202465109

Fig. 5.

Fig. 5.

mir-25 and -32 target TSC1. (A) TSC1 3′ UTR contains one predicted miR-25 and -32 binding site. The reporter assays were performed three times with essentially identical results. (B) The levels of TSC1 proteins in U87 cells at 48 h after transfection with miR-25 and -32 and scrambled oligonucleotides. (C) The levels of TSC1 protein in cells transfected with antisense oligonucleotides against miR-25 and -32 and scrambled oligonucleotides. (D) TSC1 expression levels at 48 h after knockdown of E2F1 or MYC in U87 cells, respectively. (E) The levels of TSC1 protein and mTOR activity (p-S6) in response to p53 induction in U87 cells. (F) TSC1 protein levels and mTOR activity in U87 cells treated with Nutlin-3a and transfected with miR-25 and -32. (G) Effects of miR-25 and -32 on mTOR and Akt activities at 24 h after transfection with miR-25 and -32 in U87 cells. (H) Fifty-three protein levels at 24 h after knockdown of TSC1. (I) mTOR and Akt activities in TSC1-silenced cells (U87). (A, D, and F) Data are presented as mean ± SD. We performed three biological experiments in triplicate.