Fig. 5: APR2 C-terminal region could localize at APR.
a Diagram of truncation of N-terminal fragment (2–600 aa) in endogenous APR2 in the parental parasite apr2::gfp, generating the APR2-ΔN strain. b Fluorescence of GFP in the apr2::gfp and APR2-ΔN ookinetes. Scale bars: 5 μm. Two independently performed experiments with similar results. c Quantification of GFP signal area in b. Boxes show medians with interquartile ranges, whiskers: min to max show all points. n = 28 cells examined in both groups. ****P = 3e−24, two-sided Mann–Whitney test. d Diagram and confirmation of episomal expression (EE) for HA-tagged APR2-M (501–1100 aa) and APR2-C (1001–1394 aa). Immunoblot confirmed the APR2-M and APR2-C expressed in the wildtype ookinetes (1.0 × 106 ookinetes were lysed in each sample). BiP is a loading control. Two independently performed experiments with similar results. e IFA of HA-tagged APR2-M and APR2-C episomally expressed in 17XNL and Δapr2 ookinetes. Scale bars: 5 μm. f Quantification of APR2-C IFA signal area in e. Boxes show medians with interquartile ranges, whiskers: min to max show all points. n = 22 cells examined in both groups. ****P = 1e−20, two-sided Mann–Whitney test. g Diagram of a ribosome-skipping T2A peptide insertion into the endogenous APR2 protein in 17XNL parasite, generating the APR2-T2A strain with separated expression of the N- (HA-tagged) and C- (Myc-tagged) parts of APR2. Two independently performed experiments with similar results. h Immunoblot of N- and C- parts of APR2 in the APR2-T2A ookinetes (1.0 × 106 ookinetes were lysed in each sample). P28 is a loading control. i IFA of the HA-tagged N-part and the Myc-tagged C-part of APR2 in the APR2-T2A ookinetes from 3- and 12-h in vitro culture. Merged signals were shown in white box, Scale bars: 5 μm. Two independently performed experiments with similar results. j Summary of APR2-N, APR2-M, and APR2-C in MT-binding and APR targeting.
