Fig. 1. H3K18 methylation marks parasite nuclei.

a Immunofluorescence analysis of BL3 (uninfected) mixed with TBL3 (infected) cells. Host and parasite nuclei are stained with DAPI (grey). Histone marks were detected with specific antibodies for H3K18me1 (red) or H3K18ac (cyan). Solid white arrowheads indicate the nuclei of the bovine cells and the smaller empty arrowheads point to parasite nuclei in TBL3 cells only. The yellow dotted arrows indicate the cross-section planes used for quantification. Scale bar = 5 μm. b Quantification of immunofluorescence intensity of H3K18me1 (red) and H3K18ac (blue) compared to DNA/DAPI (grey). The plot profiles are representative of the yellow cross-section lines. c Immunofluorescence staining for H3K18me1 (red) and H3K18ac (cyan) in parasite-infected macrophages (TaC12 cells). Host and parasite nuclei are stained with DAPI (grey). Leica microscope, ×100, Scale bar = 5 μm. Solid white arrowheads show the bovine host nuclei and empty arrowheads point to parasitic nuclei. The yellow dotted arrows indicate the cross-section planes used for quantification. d Quantification of immunofluorescence intensity of H3K18me1 (red) and H3K18ac (blue) compared to DNA (grey). The plot profiles are representative of the yellow cross-section lines. e Western blot analysis of BL3 and TBL3 cells (treated with the theilericidal drug Buparvaquone, Bup). H3 was used as a loading control. The bands were quantified and compared with uninfected BL3 cells. Results are representative of at least three independent experiments. f Western blot analysis of H3K18 modifications in infected TaC12 macrophages treated with or without Buparvaquone (Bup). The bands were quantified and compared with uninfected BL3 cells. Results are representative of at least three independent experiments. All these experiments were performed three times independently with similar results; representative experiments are shown. Full scans blot and immunofluorescence replicates are included in the Source Data file.