DR5-AS knockdown modulates proliferation and cell cycle in HeLa cells. (A) The rate of early apoptosis in HeLa cells under various transfection conditions was determined by flow cytometry. HeLa cells were transfected with DR5-AS GapmeR alone (DR5-AS GapmeR-1 and DR5-AS GapmeR-2) or in combination of DR5-AS GapmeR-1 with pcDNA3.1-DR5-AS (co-transfection). Only cell (no transfection), transfection reagent (no GapmeR or plasmid) and negative GapmeR were used as controls. 72 h post-transfection, the rate of early apoptotic cells was determined by Annexin V/7AAD staining. The stained cells were analyzed by a flow cytometer. (B) Proliferation rate of HeLa cells transfected with DR5-AS GapmeR alone (DR5-AS GapmeR-1 and DR5-AS GapmeR-2) or in combination of DR5-AS GapmeR-1 with pcDNA3.1-DR5-AS (cotransfection) for 72 h. Untransfected (Negative control) and transfected (Transfection Reagent, Negative GapmeR, DR5-AS-GapmeR-1 or co-transfection) HeLa cells described in Figure 4C were also subjected to cell cycle analysis and the percent of cells in each phase (C) was calculated from DNA histograms (D). (E) qPCR analyses of genes associated with cell cycle in DR5-AS knockdown HeLa cells (Negative GapmeR versus DR5-AS-GapmeR-1). ns, non-significant (p > 0.05), *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.
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