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. 2015 Mar 21;6(12):9877–9886. doi: 10.18632/oncotarget.3185

Figure 2. MORC2 phosphorylation at Ser-677 is dependent on PAK1.

Figure 2

(A) Serum treatment resulted in an increase in phosphorylation levels of endogenous MORC2 in BGC-823 cells via PAK1. BGC-823 cells were starved over night and stimulated with 10% (v/v) serum for 45 min to activate PAK1. The lysates were probed with indicated antibodies. (B) Activated Cdc42 resulted in up-regulation of MORC2 phosphorylation at Ser 677 via PAK1. BGC-823 cells were transfected with His-Cdc42Q61L and performed by Western blot. The lysates were probed with indicated antibodies. MORC2 phosphorylation at Ser677 was blotted with MORC2 phospho-specific antibody. (C) Western blot analysis of the level of MORC2 phosphorylation by blocking the upstream PAK1 expression with two different siRNAs (#1 and #2) targeting PAK1 when MORC2 was over-expressed. The lysates were probed with indicated antibodies. (D) The effect of PAK1 on MORC2 phosphorylation with ectopic MORC2 by Flag IP assays when endogenous PAK1 was knocked down. The western blot analysis was probed with indicated antibodies. (E) The effect of PAK1 on MORC2 phosphorylation with ectopic MORC2 by Flag IP assays when PAK1 was overexpressed in the presence of Cdc42 (Cdc42Q61L). The western blot analysis was probed with indicated antibodies.