ANI-2 is required for rachis bridge stabilization. (A and C) Mid-section confocal images of the germline of a wild-type (A) and an ani-2(−) (C) L4 hermaphrodite expressing a membrane marker (red) and GFP::ANI-1 (green). Bar, 10 µm. The regions delineated by the white dashed square are magnified in the inset (bar for insets, 5 µm). In A, the white arrowhead points to the germ cell opening to the rachis. (B and D) Measured fluorescence intensities (in arbitrary units specific to each curve) for each fluorescent marker along the lateral and apical cortices (white dotted line, as shown in insets; bar for insets, 5 µm) of the germ cell magnified in A and C, respectively. Red and green arrows point to peaks of membrane marker and GFP::ANI-1 fluorescence intensities, respectively, and the black arrowhead points to the intensity minimum. (E) Proportion of germ cells showing rachis bridges with a diameter >0.8 µm (turquoise) or <0.8 µm (red) in wild-type and ani-2(−) mutant animals at the L3 and L4 larval stages, as measured by fluorescent marker distribution. (F) Maximal rachis bridge diameter in germ cells of wild-type and ani-2(−) animals at the L3 and L4 larval stages, as measured with membrane (red) or GFP::ANI-1 (green) fluorescence distribution. Rachis bridges that are <0.8 µm in diameter are excluded from this analysis. Error bars represent SD. In E and F the numbers in brackets represent the total number of germ cells analyzed.