Fig. 7: Rv2067c modulates DOT1L expression. | Nature Communications

Fig. 7: Rv2067c modulates DOT1L expression.

From: The Mycobacterium tuberculosis methyltransferase Rv2067c manipulates host epigenetic programming to promote its own survival

Fig. 7

a Immunoblot depicts DOT1L levels in THP-1 cell lysates 4 and 24 h.p.i 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, for each time point. β-Actin was used as loading control. Values above the blot represent quantitation (arbitrary units). b Relative expression of DOT1L in THP-1 4 (dark gray bars) and 24 (light gray bars) h.p.i with ΔRv2067c, WtMtb and ΔRv2067c:comp normalized with respect to uninfected THP-1. Levels were normalized against GAPDH. n = 2 independent infections; p = 0.02, 0.04. 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; *P-value < 0.05. Comp stands for complemented c H3K79me3 mark in cell lysates of HEK293T with the following treatment: Lane 1: scramble shRNA (control); Lane 2: DOT1L inhibition by shRNA and Lane 3: DOT1L inhibition by shRNA along with expression of Rv2067c. H3 was used as loading control. Values above the blot represent quantitation (arbitrary units). d Graph depicts inhibition of DOT1L with small molecule inhibitor EPZ004777 which does not inhibit Rv2067c. Filled circles and squares represent scintillation counts for DOT1L and Rv2067c with EPZ004777 inhibitor at indicated concentrations. Each data point represents the mean of three replicates at each specified concentration of the compound, and the error bars represent SD. e Immunoblot analysis of H3K79me3 mark in uninfected THP-1 (left panel), inhibitor-treated THP-1 infected with M.smeg:FLAG (middle panel) and M.smeg:Rv2067c-FLAG (last panel) at indicated concentrations. H3 was kept as loading control. Source data are provided as a Source Data file.

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