Fig. 3
From: Resveratrol induces H3 and H4K16 deacetylation and H2A.X phosphorylation in Toxoplasma gondii
Effect of intracellular exposure to resveratrol (RSV) on T. gondii H2A.X phosphorylation (γH2AX). a Western blot of T. gondii recombinant H2A.X (rH2A.X; 200 ng/lane) expressed in Escherichia coli and purified by nickel resin. Rabbit anti-rTgH2A.X (α-TgH2A.X, 1:5000) and anti-T. gondii phosphorylated peptide (α-TgγH2A.X, 1:100). The phosphorylated peptide sequence was NH2- C + GKHGV-S(PO3H2)-QEF -COOH. b Western blot of T. gondii lysates with anti-SAG1 (T. gondii surface antigen 1, 1:500), anti-TgH2A.X or anti-TgγH2A.X. Lysates were obtained from purified intracellular tachyzoites previously treated with RSV (50 μM) or DMSO 0.5% for 24 h. H2A.X (arrow) and γH2A.X band intensities were quantified and normalized to SAG1 band intensities. The relative intensity of the bands (γH2A.X/H2A.X) was then calculated for each treatment. The signal normalized to SAG1 was calculated from relative values in comparison to the DMSO control. The results are the means of three replicates ± SD. This panel corresponds to Experiment 3 in Additional file 2. c Quantitation of the fold increase in γH2A.X level after RSV treatment. d Putative model of RSV action that leads to a decrease in T. gondii replication. Dotted lines indicate reduced activities or processes. For acetylations and phosphorylations, color intensity represents the PTM mark level