Fig. 8: Downstream events consequent to H3K79 methylation by Rv2067c.

a Fold enrichment of H3K79me3 at specific loci (as indicated) added by Rv2067c in THP-1 infected with ΔRv2067c (dark gray), WtMtb (light gray) and ΔRv2067c:comp (checker). Data is shown as fold enrichment over IgG control. b Relative expression of TMTC1, NLRC3 and SESTRIN3 in THP-1 infected with ΔRv2067c (dark gray), WtMtb (light gray) and ΔRv2067c:comp (checker) with respect to uninfected THP-1 macrophages. Ct values were normalized against GAPDH. c Graph shows relative expression of indicated cytokines in THP-1 infected with ΔRv2067c (dark gray), WtMtb (light gray) and ΔRv2067c:comp (checker). Ct values were normalized against GAPDH. d Fold enrichment of H3K79me3 mark at the promoter region of IL-6 and TNF-α in THP-1 infected with ΔRv2067c (dark gray), WtMtb (light gray) and ΔRv2067c:comp (checker). Data is shown as fold enrichment over IgG control. a–d For all ChIP and qRT-PCR, n = 2 independent infections. Data is plotted as mean and error bars represent SD. P-values depicted on the graphs were calculated using unpaired two-tailed Student’s t-test. e, f Immunoblots depict levels of caspase-8, necrotic, pro- and anti-apoptotic markers as indicated in THP-1 cell lysates infected with various Mtb strains. Lane 1: uninfected THP-1; Lane 2, 3 and 4: THP-1 infected with ΔRv2067c, WtMtb and ΔRv2067c:comp strains, respectively. β-Actin was used as loading control for TNF- α and caspase-8 and H3 as loading control for RIPK1, FADD and RIPK3 (e). H3 was used as loading control (f). Values above the blot represent quantitation (arbitrary units). g Graph depicts the percentage of apoptotic and necrotic macrophages analyzed by flow cytometry upon infection with ΔRv2067c (dark gray), WtMtb (light gray) and ΔRv2067c:comp (checker) n = 2 independent infections. Data is plotted as mean and error bars represent SD. P-values depicted on the graphs were calculated using unpaired two-tailed Student’s t-test. Source data are provided as a Source Data file.