Fig. 4. Western blot analysis of A. franciscana B-subunit fusion protein (F1, 4 µg NtA column purified; F2, 7 µg solublized inclusion body), V-ATPase expression in subcellular fractions of encysted embryos (M, mitochondrial pellet; H, heavy membrane pellet; V¹⁰⁰, heavy microsomal vesicle pellet; S¹⁰⁰, supernatant of 100 000 g spin; 40 µg per lane) and a Manduca sexta positive control (Ms, heavy membrane pellet; 1 µg per lane). The blot on the left was incubated with primary antibody raised against the native V-ATPase V₁ complex purified from Manduca sexta midgut (Huss, 2001). All immunoreactive bands were identified by comparison with control blots incubated with secondary antibody only (blot not shown) and are listed in parentheses (together with their apparent M_r in kDa) to the left of the figure. The blot on the right (lane CM) is the control for mitochondrial lysates from A. franciscana (incubated with secondary antibody only) and is included to demonstrate that the banding in lane M is nonspecific.