FIG 2.

CWIP is activated during asexual development and is accompanied by RlmA phosphorylation. (A and B) Western blotting assay of MpkA phosphorylation in a time course experiment of synchronized asexual differentiation. α-(P)-p44/42 or α-p44/42 antibodies were used to detect the phosphorylation and total MpkA, respectively. α-γ-Tubulin antibody was used as a loading control. Signal intensities were quantified, and the ratios of (P)-MpkA to MpkA were calculated and are shown below the panels. (C) Western blot of protein from the rlmA::3×FLAG treated (+ CIP) or not (− CIP) with calf intestinal alkaline phosphatase and probed with α-FLAG antibody in membranes obtained from Phos-tag or regular 8% SDS-PAGE. The arrows point to the phosphorylated forms of RlmA, and the arrowhead points to the unphosphorylated protein. Predicted protein sizes on blot: MpkA, 48.5 kDa; and RlmA::3×FLAG, 70.3 kDa.