Figure 2
PTEN-sh is processed by micro-processor Drosha/DGCR8 and Dicer and functions at the post-transcriptional level. (A) The effect of PTEN-sh on the luciferase activities of pGL3-PTEN-3′UTR was examined with a luciferase reporter assay in 293T cells. Vector indicates empty plasmid DNA. (B) The effect of PTEN-sh and PTEN-sh-mut on the expression of PTEN and p-Akt level was assessed with a Western blot analysis in L-O2 cells. (C) The effects of Drosha and DGCR8 on luciferase activities of pGL3-PTEN-3′UTR were measured with a luciferase reporter assay in 293T cells. (D) PTEN mRNA levels were examined by RT-PCR in L-O2 cells when Drosha and DGCR8 were knocked down by siRNA. (E) The interaction between PTEN mRNA and Drosha or DGCR8 was examined with an RIP analysis. (F) The effects of 100 nmol/L siDicer on the luciferase activities of pGL3-PTEN-3′UTR induced by PTEN-sh were detected in 293T cells. Mean±SD. n=3. Each experiment was repeated three times. *P<0.05, **P<0.01 vs vector. NSP>0.05, #P<0.05, ##P<0.01 vs siNC, unpaired Student's two-tailed t-test.
