Figure 3. MEL-26 Localizes to the Cleavage Furrow and Accumulates at the Cell Cortex in
cul-3(RNAi) and
rfl-1(
or198) Embryos. (A and C) MEL-26 was localized in wild-type and the indicated mutant embryos by indirect immunofluorescence using affinity-purified MEL-26 antibodies and photographed by fluorescence microscopy. In
cul-3(RNAi) or
rfl-1(RNAi)
rfl-1(
or198) embryos, MEL-26 also accumulated at the cortex (arrows). The centrosomal staining occasionally observed with anti-MEL-26 antibodies is not specific, as it was also observed in embryos depleted for MEL-26 by RNAi. The DNA was stained by Hoechst as described in the Experimental Procedures. The arrowheads in (C) point to tips of ingressing cleavage furrows during mitosis (top and bottom panels) and ruffles during pronuclear migration (middle panel). (B) The accumulation of MEL-26 was analyzed in wild-type,
rfl-1(
or198), and
mel-26(RNAi) embryo extracts by immunoblotting with MEL-26 antibodies (upper panel); immunoblotting with antibodies against tubulin served as a loading control (lower panel).