FIGURE 7:

TORC2 and RasC responses to folate and cAMP are nonadditive. (A) AKT and PKBR1 phosphorylations are nonadditive in response to a mixture of saturated cAMP and folate. Cells were stimulated either with 10 μM cAMP plus 50 μM folate or 10 μM cAMP plus 50 μM folate. TORC2 phosphorylation of AKT and PKBR1 was assayed by immunoblot at the times indicated. For quantification, see Supplemental Figure S6A. (B) RasC activation is nonadditive in response to a mixture of saturated cAMP and folate. FLAG-RasC–expressing cells were stimulated either with 50 μM folate plus 10 μM cAMP, or 50 μM folate plus 10 μM cAMP. RasC-GTP levels were determined by interaction-specific affinity and normalized to total RasC by α-FLAG immunoblot assay at the times indicated. For quantification, see Supplemental Figure S6B.