FIG 3 .

Apicoplast protein import and membrane expansion are not affected in the first cycle of ATG8 knockdown. (A) Processing of a luminal apicoplast protein, ClpP, in ATG8⁺ or ATG8-deficient parasites approximately 24 h after aTC removal. Apicoplast-minus parasites generated by chloramphenicol treatment and IPP rescue over 4 replication cycles (25), which possess only precursor ClpP, are shown for reference. preClpP, full-length (precursor) form of ClpP, 43 kDa; mClpP, mature (apicoplast-luminal) ClpP, 25 kDa. The asterisk indicates a nonspecific band. ATG8 expression in the corresponding time points is shown for reference. Numbers on the left are molecular masses in kilodaltons. (B) Quantification of parasites with the indicated apicoplast morphology during the first cycle of ATG8 knockdown, 32 h after aTC removal. Apicoplast was visualized using the luminal apicoplast marker ACP_L-GFP. Mean ± standard deviation of 2 independent experiments is shown. The P value was determined by unpaired two-tailed t test for the “tubulated” category. Representative images and data for separate replicates are shown in Fig. S3. ns, not significant.