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. 2017 Jan 5;5:e2859. doi: 10.7717/peerj.2859

Figure 5. Expression change of PTEN/PI3K/Akt pathway in the process of miR-21 mimics induced proliferation in c-kit+ CSCs.

Figure 5

Cultured CSCs were treated with miR-21 mimics for 48 h before the subsequent procedures. To test the contribution of PTEN/PI3K/Akt signaling to miR-21 mimics’s pro-proliferation effects in c-kit+ CSCs, PTEN and PI3K were inhibited with Phen or LY294002 respectively. (A) RT-PCR was carried out to detect miR-21 mimics’s effects on PTEN expression at the mRNA level, which showed no change between Control, miR-21 scramble, miR-21 mimics and miR-21 mimics+ LY294002 group, while Phen resulted in a significant down-regulation of PTEN compared with the other groups. (B–C) Western blot was carried out to detect miR-21 mimics’s effects on PTEN protein expression, which showed that miR-21 mimics significantly down-regulated PTEN protein in miR-21 mimics group compared with the scramble group. In addition, both Phen treatment and miR-21 mimics incubation increased p-Akt level, while PI3K inhibitor LY294002 decreased p-Akt level dramatically (P < 0.05). a, P < 0.05 compared with Control; b, P < 0.05 compared with miR-21 scramble group; c, P < 0.05 compared with miR-21 mimics group; d, P < 0.05 compared with Phen group. n = 3 in each group. p-Akt = phosphor-Akt.