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. Author manuscript; available in PMC: 2018 Nov 16.
Published in final edited form as: Biochem J. 2017 Nov 16;474(23):3903–3914. doi: 10.1042/BCJ20170279

Figure 4. CA-Rac1-mediated activation of PI3Kβ requires Gβγ.

Figure 4

(A) Stable p110β knockdown cells were transiently transfected with wild-type p110β, p110βKK-DD or p110βRBD, with or without GFP-CA-Rac1. After 48 h, cells were lysed and blotted for p110β, Akt, and pT308-Akt. (B) Quantitation of pAkt/Akt immunoblots in (A). pAkt/Akt ratios in each experiment were normalized to that seen in cells expressing wild-type p110β and GFP-CA-Rac1. The data represent the mean ± SEM from three independent experiments. (C) Parental MDA-MB-231 cells were transfected with GFP-CA-Rac1 with or without co-transfection of GRKct-PM. Representative immunoblots show pT308-Akt, total Akt, GFP-CA-Rac1, and GRKct-PM. (D) Quantitation of pAkt/Akt levels from immunoblots in (C). pAkt/Akt ratios in each experiment were normalized to that seen in cells expressing GFP-CA-Rac1 without GRKct-PM. The data represent the mean ± SEM from five independent experiments. (E) MDA-MB-231 cells were transfected with GFP-CA-Rac1 with or without PTX treatment. Representative immunoblots show pT308-Akt, total Akt, and GFP-CA-Rac1. (F) Quantitation of pAkt/Akt levels from immunoblots in (E). pAkt/Akt ratios in each experiment were normalized to that seen in cells transfected with GFP-CA-Rac1 without PTX treatment. The data represent the mean ± SEM from three independent experiments. *P < 0.05; ****P < 0.0001.