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. 2021 May 31;118(23):e2011278118. doi: 10.1073/pnas.2011278118

Fig. 2.

Fig. 2.

SETD2-mediated H3K14me3 levels increase in response to replication stress. (A) Histone extracts from HeLa cells were treated with 40 μM etoposide (VP-16) for 4 h, 1 mM DOX for 8 h, or 6 mM HU for 3 h. “CTR” indicates the cells without any treatment. (B) HeLa cells were treated with the indicated concentrations of HU for 5 h. The chromatin fractions were extracted for Western blotting. “CTR” indicates the cells without any treatment. (C) HeLa cells were treated with 2 mM HU for the indicated times. The chromatin fractions were extracted for Western blotting. “CTR” indicates the cells without any treatment. (D) HeLa cells were treated with 4 mM HU for 5 h. The chromatin fractions were extracted for Western blotting. “CTR” indicates the cells without any treatment. (E) HeLa parental cells and SETD2-KO HeLa cells were treated with or without 4 mM HU treatment for 5 h. The chromatin fractions were extracted for Western blotting. “CTR” indicates the cells without any treatment. (F) A statistical analysis of E was performed by scanning the density of H3K14me3 and RPA32 pS33 band of the Western blots. The band density of HeLa parental cells in “CTR” was normalized to 1. Data are shown as means ± SD (n = 3). (G) HeLa cells transfected with SUV39H1 siRNAs or a nonspecific siRNA (NC) were treated with or without 4 mM HU treatment for 5 h. The chromatin fractions were extracted for Western blotting. “CTR” indicates the cells without any treatment. (H) A statistical analysis of G was performed by scanning the density of H3K14me3 and RPA32 pS33 band of the Western blots. The band density of HeLa cells transfected with “NC” siRNAs in “CTR” was normalized to 1. Data are shown as means ± SD (n = 3).