Generation of DPAP3 cKO lines. D. IFA analysis of mature schizonts showing the loss of DPAP3 signal after RAP (rapamycin) treatment. Ring-stage F8cKO parasites were treated with DMSO or RAP for 3 h and fixed for IFA analysis at 48 h.p.i. Slides were stained with anti-mCherry (green) and anti-MSP1 (red) or anti-SUB1 (red) antibodies. DNA was stained with DAPI (blue); scale bar: 5μm. Lower panel: IFA analysis of the complementation lines showing colocalization of chromosomal DPAP3-mCh and episomal DPAP3-HA expressed under that dpap3 or ama1 promoters. Mature schizonts from the F8cKO+WTdpap3, F8cKO+MUTdpap3, F8cKO+WTama1, and F8cKO +MUTama1 were fixed and stained with anti-HA (red) and anti-mCherry (green). DNA was stained with DAPI (blue); scale bar: 5μm.

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