Fig. 3: TgFLP12 disruption is detrimental to tachyzoite development and apicoplast maintenance.

a Inducible knockdown of TgFLP12 in the TgFLP12-HA line after promoter replacement. TgFLP12 was detected by Western Blot using Rt anti-HA antibody, and complete shutdown of the protein occurred after 48 h of ATc treatment. TOM40 (lower panel) served as loading control. Illustrative WB from at least three biological replicates. b Detrimental effect of TgFLP12 loss shown by plaque assays performed in the absence (-) or presence (+) of ATc and with different FBS concentrations (0, 1 and 10%) and fixed after 7 days. c Plaque size quantification and statistic from plaque assay (b). Data are presented as mean values +/− SD (n = 3). d The absence of TgFLP12 reduced the intracellular replication of Toxoplasma tachyzoites. Two-way Anova, Šídák’s multiple comparisons, data are presented as mean values +/− SD (n = 4.) e Apicoplast disruption in absence of TgFLP12 revealed by IFA and using an anti-CPN60 antibody, Blue = Hoechst, Red = apicoplast marker CPN60, and green = 3xHA tag. f Statistical analysis of the apicoplast disruption after complete shutdown of TgFLP12 expression. Data are presented as mean values +/− SD (n = 3). g Transmission electron microscopy showing apicoplast morphology after 48 h of ATc treatment on TgFLP12-iKD, apicoplast appeared swelling and accumulating electron light material in the stroma (redhead arrows) as well as in the intermembrane spaces (red arrows). Images are illustrative of average observations on at least three biological replicates.