Figure 2

Validation of anti-pSer1333 ROCKI antibody. (A) Wild-type (WT) or S1333A mutant of flag-ROCKI proteins were expressed in HEK293T cells and immunoprecipitated with anti-flag antibody followed by probed with anti-flag or anti-pSer1333 ROCKI antibody with or without peptide competition in a same set of sample. (B) WT flag-ROCKI immunoprecipitates were incubated with or without λPPase at 30°C for 20 min followed by Western blot analysis as indicated. (C) Flag-tagged ROCKI and ROCKII proteins were immunoprecipitated for Western blot analysis using anti-pSer1333 ROCKI, anti-pSer1366 ROCKII, and anti-flag antibodies in a same set of sample.