Drak and Rok both regulate Sqh phosphorylation. (A and B) Western blot analysis of 4–6 h APF pupal lysates prepared from wild-type (WT), drakKO, and drakdel pupae, using anti-phospho-MRLC, anti-phospho-Moesin, and anti-Sqh antibodies. Levels of phospho-Sqh, normalized to total Sqh levels, are plotted in (B). (C and D) Western blot analysis of dorsal closure-stage wild-type, drakdel, rok2, and drakdel,rok2 double mutant embryo lysates, using anti-phospho-MRLC, anti-phospho-Moesin, and anti-Sqh antibodies. Levels of phospho-Sqh, normalized to total Sqh levels, are plotted in (D). (E and F) Western blot analysis of third instar wing imaginal disc lysates prepared from wild-type, drakdel/drakKO, rok2/+, and drakdel,rok2/drakKO larvae, using anti-phospho-MRLC and anti-Sqh antibodies. Levels of phospho-Sqh, normalized to total Sqh levels, are plotted in (F).