Extended Data Fig. 4: Orf8 effects on histone PTMs.
From: SARS-CoV-2 disrupts host epigenetic regulation via histone mimicry
(a) Orf8 does not significantly affect H3K27ac. N = 616 (GFP), 550 (Orf8) from 3 independent transfections. (b) H3 serine 10 phosphorylation (H3S10ph), H3K9 dimethylation (H3K9me2), Lamin B and global acetylation (of histones or of non-histone proteins) in GFP or Orf8 expressing cells isolated by FACS. (c) Orf8 constructs tested show similar levels of expression in HEK293T cells. N = 137 (Orf8), 87 (Orf8ΔARKSAP), 120 (S84L) from 4 independent transfections. (d–f) Orf8 with a mutation commonly found in the SARS-CoV-2 genome, Orf8-S84L, shows the same effects on histone PTMs H3K9me3 (d), H3K27me3 (e), and H3K9ac (f). N = 332 (GFP), 237 (S84L) cells for H3K9me3; 186, 166 cells for H3K27me3; 332, 237 cells for H3K9ac from 2 independent transfections. (g) Orf8 with a 6 amino acid deletion outside of the ARKSAP motif shows the same effects on H3K9me3. N = 216 (GFP), 120 (Orf8), 88 (AGSKSP) cells from 2 independent transfections. (a,g) 1-way ANOVA with post-hoc 2-sided t-test and Bonferroni correction. (b,d,e,f) 2-sided t-test. Scale bar = 10μM. For gel source data, see Supplementary Fig. 1n-m.
