Fine mapping of MIC5 propeptide confirmed that an N-terminal leucine is important for trafficking. A. Schematic representation and sequences of full-length, deletion and replacement mutants of MIC5 propeptide. An arrow denotes the proteolytic cleavage site. B. Location of mature MIC5 in intracellular parasites was examined by IFA using anti-MIC5 and anti-MIC10 antibodies. Arrows denote the sites where the mature MIC5 is retained in intracellular parasites that showed defective trafficking. Scale bar: 2 μM. C. Quantification of MIC5 and MIC10 colocalization. Bars represent SEM. ns, not statistically different from WT i.e., P>0.05, one sample t-test. *, P<0.05. D. Parasites stably transfected with full-length propeptide or deletion mutants were analyzed by immunoblotting with anti-MIC5.