Fig. 3: Effect of PDE4DIP knockdown on apoptosis and cell cycle progression.

A KEGG enrichment analysis of the pathways affected by knockdown of PDE4DIP in A549 cells. B PDE4DIP-KD-induced apoptosis was analysed via flow cytometry. Cells were infected with nontargeting control shRNA (shNC) or PDE4DIP shRNA (shP1 or shP2) for 84 h. Left panel, representative images of the apoptosis assay results; right panel, calculated numbers of apoptotic cells (%, Q4). C Apoptosis was examined by a TUNEL assay in PDE4DIP-KD NSCLC tissues from xenograft. Upper panel, representative images of TUNEL positivity, with the arrow indicating TUNEL-positive cells; lower panel, histograms showing the quantification of TUNEL-positive cells. Scale bar: 100 μm. D Flow cytometric analysis of the cell cycle in PDE4DIP-KD NSCLC cells. Cells were infected with lentivirus containing shP1, shP2 or the nontargeting shNC for 84 h. Upper panel, representative images of the cell cycle analysis results; lower panel, histograms showing the quantification of the cell cycle distribution. E Western blot analysis of apoptosis-related proteins in control and PDE4DIP-KD cells. Cl Casp-3/9, cleaved caspase-3/9. F Silencing Bim abolished PDE4DIP-KD-induced apoptosis. Left panel, evaluation of interference with Bim expression by Western blotting; right panel, histograms showing the quantification of apoptosis. G Western blot analysis of cell cycle-related proteins in control and PDE4DIP-KD cells (B–D and F). The data are shown as the means ± SD of a representative experiment performed in triplicate. **P < 0.01; ***P < 0.001 vs. shNC.