Extended Data Fig. 2: H4S47 O-GlcNAcylation influences cell proliferation.
From: H4S47 O-GlcNAcylation regulates the activation of mammalian replication origins
a-c, Impact from H4S47 O-GlcNAcylation on cell proliferation. HEK293T (a), HT1080 (b) and A549 (c) cells were transfected with Flag-H4WT or -H4S47A and incubated with or without PUG (100 μM). Cell numbers were counted every 24 h. n = 3 biologically independent experiments. Two-way ANOVA Tukey’s multiple comparisons test, means ± s.d., n.s., non-significant. d, Functional assessment of mutations at H4S47 in DNA replication. HEK293T cells were transfected with Flag-H4WT, Flag-H4S47A, Flag-H4S47D or Flag-H4S47E for 48 h prior to the detection of replication efficiency by FACS analysis. e, DNA fiber assay for replication fork function. Frequency distribution of CIdU track lengths (left) and inter-origin distances (right) for experiments in Fig. 1d were shown. f, Assessment of DNA damage response. HEK293T cells transfected and treated as in Fig. 1a were analyzed by WB with indicated antibodies. HU (3 mM) treatment for 3 h was included as a positive control.
