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. 2011 Dec 21;287(9):6469–6481. doi: 10.1074/jbc.M111.324582

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Transient HDHB depletion does not disrupt replication stress-induced checkpoint signaling but impairs recovery of HeLa cells from replication stress. A, whole cell extracts of HeLa cells transiently expressing non-silencing (Ctl) or HDHB-silencing shRNAs (H1 and H2) were analyzed by Western blotting with the indicated antibodies. B, HeLa cells transiently expressing non-silencing (Ctl) or HDHB-silencing shRNAs (H1 and H2) were incubated with 10 μm BrdU for 30 min and stained for BrdU and total DNA and analyzed by flow cytometry. C, HeLa cells transiently expressing non-silencing (Ctl) or HDHB-silencing shRNAs (H1 and H2) as in A were exposed to 2 mm HU for 2 h or left untreated (−) as indicated. Whole cell extracts were then analyzed by SDS-PAGE and Western blotting with antibodies against HDHB, Chk1 phospho-Ser-317, RPA32 phospho-Ser-4, Ser-8, total RPA32 (RPA2), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a loading control.