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. 2017 Sep 22;13(9):e1006615. doi: 10.1371/journal.ppat.1006615

Fig 1. HAT2 is constitutively nuclear and acetylates H4K10 in vitro.

Fig 1

a. Schematic depiction of conserved motifs and residues in the Leishmania donovani HAT2 gene. MOZ/SAS: conserved core catalytic acetyltransferase domain. Cys273 and Glu332: the conserved catalytically important cysteine and glutamate residues. b. Left panel: Western blot analysis of immunoprecipitation reaction from transfectant cells expressing the tagged HAT2-eGFP, performed using anti-eGFP antibodies (1:2000 dilution; 6.75x108 cells used per IP reaction). Right panels: IFA of HAT2-eGFP at different cell cycle stages. DAPI: stains DNA compartments; N: nucleus, K: kinetoplast. G1/early S: one nucleus, one short kinetoplast (1N1K); late S/early G2/M: one nucleus, one elongated kinetoplast (1N1K); late G2/M: two nucleii, one kinetoplast (2N1K); post-mitosis—two nucleii, two kinetoplasts (2N2K). Imaging done using a LeicaTCS SP5 confocal microscope, 100X (in oil) objective; analysis using Leica LAS AF software. Magnification bar: 5 μm. c.—e. HAT assays performed with various peptide substrates (sequences in boxes above graphs). Auto: no peptide added. Inset (first panel): western blot analysis of 1/6 pulled-down fraction.