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. 2009 Jan 1;20(1):1–9. doi: 10.1091/mbc.E08-01-0036

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© 2008 by The American Society for Cell Biology

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Figure 3. — UbcH7 knockdown delays progression from S to G2 phase. HeLa cells were treated for 48 h with siRNA as indicated. Cells were then treated overnight with 2 mM HU for synchronization. (A) Immunoblot showing knockdown of UbcH7. (B) Cell cycle profile of cells at times after drug release. Cell cycle numbers are from duplicate samples. Similar delays in S phase progression to G2 were also observed with another UbcH7 siRNA sequence. (C) Top, cyclin A and E1 levels from samples treated with NS siRNA or UbcH7-specific siRNA immediately after drug release or 12 h after release. Bottom panel, quantitation of cyclin A levels normalized to E1 levels. Normalized cyclin A levels were compared between UbcH7 siRNA-treated cells and NS siRNA-treated cells at each time point. (D) Knockdown of UbcH7 decreases cell proliferation. Comparison of MTS OD between NS siRNA and UbcH7 siRNA at 72 and 96 h of knockdown.