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. 2012 May;14(5):440–450. doi: 10.1593/neo.12352

Figure 4.

Figure 4

BART has GAP activity toward Rac1. (A) The BART-rescue construct was transfected into control and BART knockdown cells of S2-013, and 48 hours later, the amount of active GTP-loaded Rac1 was determined by GST-pull-down assays using GST-PAK-CRIB. Precipitates were examined by Western blot analysis using an anti-Rac1 antibody. Data are representative of three independent experiments. Closed arrowhead indicates exogenous BART; open arrowhead, endogenous BART. (B) The possibility that BART might have a GAP function for Rac1 GTPase was assayed by in vitro GAP assays using human Rac1 with or without the human p50 RhoGAP protein together with recombinant BART protein. GST was used as a negative control for RhoGAP and BART protein. GAP activity was assayed by measurement of the phosphate generated by hydrolysis of GTP. Data are representative of three independent experiments and are shown as means ± SEM. *P < .005 compared with their respective controls.