The cytoplasmic domain of STEVOR contributes to infected erythrocyte stiffness. (A) Schematic representation of STEVOR recombinant proteins overexpressed in the Full-Ty1 and in the Trunc-Ty1 lines. Signal peptide (orange, SP), PEXEL/HT motif (black bar), N-terminal semiconserved region (light blue, C1), hypervariable region (dark blue, V), transmembrane domain (yellow, TM), C-terminal conserved cytoplasmic region (light blue, C2), and Ty1-tag (green star) are indicated. (B,E) Retention rates in microsphilters of stage III GIE (B) and asexual stages (E) from the control (light gray), the Full-Ty1, the Trunc-Ty1, and the Null lines cultivated with (+BSD, dark gray) or without (−BSD, medium gray) BSD. Error bars denote the standard error of the mean. Outliers are shown as open circles. Stars represent highly significant differences in retention rates (***P < .001; **P < .01). n, number of experiments; ns, nonsignificant differences in retention rates. (C,F) Western blot analysis of STEVOR expression in stage III GIE (C) and asexual stages (F) from the Full-Ty1 and the Trunc-Ty1 lines cultivated with (+BSD) or without (−BSD) BSD. Immunoblots were probed with a rabbit polyclonal antibody directed against STEVOR (PFC0025c) and with a rat polyclonal antibody directed against HSP70 to normalize expression. Quantitation of signal intensities was realized using Quantity One software (Bio-Rad). (D,G) Immunofluorescence analysis of stage III GIE (D) and asexual stages (G) from the Full-Ty1 and the Trunc-Ty1 lines. Infected erythrocytes were stained with anti-Ty1 antibodies followed by anti-mouse Alexa 488–conjugated IgG. Pictures were taken under identical exposure conditions. The bars represent 2 µm.