Fig. 3

FR235222-mediated inhibition of HDAC3 induces BCLA in a strain-specific manner. a HFFs were infected with WT or Δbcla parasites (Pruku80 or 76K), and expression of BCLA was monitored following HDAC3 chemical inactivation by FR235222 and BCLA (red) staining. Cells were co-stained with Hoechst DNA-specific dye (blue) or GFP (green). BCLA is not labeled in Δbcla strains, therefore validating the specificity of the anti-BCLA antibodies. Scale bar, 10 μm. b Pruku80 or RHku80 strains were engineered to endogenously epitope tag with HA BCLA. Expression was monitored following HDAC3 chemical inactivation by FR235222 following by HA and Hoechst DNA-specific dye co-staining. The experiment was conducted more than three times, and representative images are displayed. Scale bars, 10 μm. c Expression of BCLA (red) following HDAC3 chemical inactivation by FR235222 was measured by IFA in HFFs infected with type I (RHΔku80 and GT1), type II (PruΔku80, 76K-GFP-luc, and ME49), and type III (CTG) strains of T. gondii. Scale bar, 10 μm. Right graph: quantification of the intensity of BCLA in each PV after FR235222 stimulation. Each symbol marks the BCLA density of a single PV. The results are represented as mean ± standard deviations from two independent experiments; the number of PVs quantified was at least 70. Statistical significance when comparing each individual FR235222-treated strain and the corresponding control (DMSO, 0.1%) was determined by a non-parametric Mann-Whitney test