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Figure 6. B55α-mediates RPA2 dephosphorylation. (A) HeLa cells with or without expression of HA-B55α, were treated with HU (1 mM, 24 h) as indicated. The cell lysates were analyzed by immunoblotting for RPA2, B55α, phospho-RPA2 Ser-4/Ser-8, Ser-33, phospho-Chk1 Ser-317, and β-actin. (B) The level of RPA2 S4/8 phosphorylation was examined by immunoblotting, as in panel A, and quantified using ImageJ. The mean values and standard deviations were calculated from three independent experiments. Statistical significance was analyzed using an unpaired 2-tailed Student’s t-test. A p-value < 0.001 was considered statistically highly significant (**). (C) HeLa cells were treated with B55α expression and HU, as in panel A. Immunofluorescence was performed using phospho-RPA2 Ser-4/Ser-8 antibody. (D) The percentage of cells exhibited positive RPA2 S4/8 phosphorylation was examined by immunofluorescence, as in panel C, and quantified. The mean values and standard deviations were calculated from three independent experiments. Statistical significance was analyzed using an unpaired 2-tailed Student’s t-test. A p-value < 0.05 was considered statistically significant (*). Image published in: Wang F et al. (2018) © The Author(s) 2018. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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