FIGURE 4:

Lgd regulates CHMP4 recruitment to MVEs. (A) Representative immunoblot using antibodies directed against Lgd (top) or β-actin (bottom) of extracts generated from control and lgd mutant animals, following separation by SDS–PAGE (n = 3). (B) Representative images taken in utero of control and lgd mutant animals expressing a GFP fusion to Cav1 using confocal microscopy are shown (n = 10 animals each). Bar, 10 μm. (C–E) Representative images of control and lgd mutant zygotes expressing a GFP fusion to Cav1 and immunostained using antibodies directed against Hrs (C), Tsg101 (D), or CHMP4 (E) are shown (n = 5 animals each; zoomed portions of embryos are shown). Bars, 2 μm. (F) Quantification showing the relative level of overlap between Cav1 and various ESCRT subunits in animals lacking Lgd as compared with control animals. *, p < 0.05, based on an ANOVA followed by a Tukey post hoc test. (G) Representative images of control and lgd mutant zygotes expressing a GFP fusion to Cav1 and immunostained using antibodies directed against CHMP4 are shown following depletion of Vps4 (n = 5 animals each; zoomed portions of embryos are shown). Bar, 2 μm.