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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Cell Microbiol. 2012 Jun 26;14(10):1544–1553. doi: 10.1111/j.1462-5822.2012.01819.x

Figure 1. Rac1 activation of HBMEC by C. neoformans.

Figure 1

A. To examine host cell activation of Rac1 by C. neoformans, HBMEC were incubated with C. neoformans serotype D strain B3501A for various time periods. Activated Rac1 (GTP-Rac1) were pulled down by incubation of HBMEC lysates with PBD-GST beads that bind GTP-Rac1 and beads examined for Rac1 in Western Blot assay. Total Rac1 in lysates was examined as a loading control. Band intensities was estimated densitometrically and then represented as a ratio of GTP-Rac1 to total Rac1 assuming Time 0 as 1.

B. The specificity of Rac1 activation was assessed by incubating HBMEC with various concentrations of NSC23766 (a specific inhibitor of Rac1) after which the host cells were incubated with strain B3501A for 30 min and then examined for GTP-Rac1.

Band intensities was estimated densitometrically and then represented as a ratio of GTP-Rac1 to total Rac1 assuming time 0 = 1.