Figure 5
The inhibitor (PD0325901) of the MAPK/ERK pathway blocked hPMSC proliferation, which was facilitated by HIF-2α.
(A) The results of the CCK-8 assay of the processed hPMSCs in the PD0325901- or DMSO-treated groups after (I) comparisons of the cumulative cell growth between hPMSCs over-expressing HIF-2α and hPMSCs-empty vector-NC in the inhibitor- or vehicle-treated groups, and (II) comparisons of the cumulative cell growth between the HIF-2α-silenced hPMSCs (shRNA1545) and hPMSCs-scramble shRNA-NC in the inhibitor- or vehicle-treated groups. The PD0325901 (inhibitor) treatment was marked as +, but the DMSO (vehicle) treatment was not marked. The tests were performed in triplicate and repeated in three independent experiments. The data are presented as the mean ± S.D. (error bars). (B) EdU flow cytometry cell proliferation assays of the hPMSCs that were treated with either PD0325901 or DMSO for 2 days. (I) comparisons of proliferation rates between hPMSCs over-expressing HIF-2α and hPMSCs-empty vector-NC in inhibitor- or vehicle-treated groups, and (II) comparisons of proliferation rates between hPMSCs silencing of HIF-2α (shRNA-1545) and hPMSCs-scramble shRNA-NC in inhibitor- or vehicle-treated groups. PD0325901-treated groups are marked with “+”; DMSO-treated groups are not marked. Tests were performed in triplicate and repeated in three independent experiments. Data results were presented as the mean ± S.D. (error bar) and were statistically analyzed by one-way ANOVA. ***p < 0.001. (C) The qRT-PCR results of the processed hPMSCs in the PD0325901- or DMSO-treated groups; (I) comparisons of CCND1 and MYC expression between the hPMSCs over-expressing HIF-2α and the hPMSCs-empty vector-NC in the inhibitor- or vehicle-treated groups, and (II) comparisons of CCND1 and MYC expression between the HIF-2α-silenced hPMSCs (shRNA1545) and hPMSCs-scramble shRNA-NC in the inhibitor- or vehicle-treated groups. The PD0325901 (inhibitor) treatment was marked as +, but the DMSO (vehicle) treatment was not marked. The tests were performed in triplicate and repeated in three independent experiments. The data are presented as the mean ± S.D. (error bars) and were statistically analyzed using one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001.
