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. Author manuscript; available in PMC: 2011 Dec 1.
Published in final edited form as: Cancer Res. 2010 Nov 23;70(23):10002–10011. doi: 10.1158/0008-5472.CAN-10-2606

Figure 6.

Figure 6

PKA activity is increased by LY294002 and LY303511 and required for enhanced GJIC. (A) Immunofluorescence analysis of CREB phosphorylated at Ser 133 in MDA-MB-231 revealed an increase in phosphorylation after 2 hr treatment with LY294002 (10 μM) and LY303511 (10 μM). (B) Analysis of phosphorylation of CREB at Ser 133 in nuclear extracts from MDA-MB-231 treated with LY294002 (10 μM) showed phosphorylation of CREB within 15 min. Treatment with the PKA agonist 8-BR-cAMP (0.1 mM, 0.5 mM, 1.0 mM) increased GJIC in a dose dependent manner in MDA-MB-231 (C), while inhibition of PKA activity by H89 (10 μM) when used in co-treatments with LY294002 (10 μM) inhibited GJIC compared to LY294002 alone (D, calcein) (E, quantified). [*=p<0.05]. Pretreatment of cells with the adenylate cyclase inhibitors 2′5′-dideoxyadenosine (F) or SQ 22,536 (G) (10 μM,75 μM,100 μM) did not reduce the ability of LY294002 (10 μM) to increase GJIC.