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. 2011 Apr 18;193(2):347–363. doi: 10.1083/jcb.201010034

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© 2011 Day et al.

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Figure 6. — AKAP11 targets free RIα. (A) Prkar1a^−/− MEFs were transiently cotransfected with GFP-tagged RIα (green) and mCherry-tagged PKAc (red) and stained for endogenous AKAP11 (blue). (B) Prkar1a^−/− MEFs were transiently cotransfected with GFP-tagged RIα (green) and mCherry-tagged PKAc (red), treated with 20 µM forskolin (FSK)/200 µM IBMX, and then stained for endogenous AKAP11 (blue). (C) siRNA of AKAP11 effectively knocked down the levels of endogenous AKAP11 protein in Prkar1a^−/− MEFs after 1–2 d. No effect on the expression level of transiently transfected RIα-GFP was observed, as judged by immunoblotting of whole-cell extracts with antibodies against AKAP11, RIα, Hsp90, or tubulin. (D) Knockdown of endogenous AKAP11 disrupted the formation of RIα-GFP puncta in Prkar1a^−/− MEFs. Ctrl, control. wt, wild type. Bars, 20 µm.