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. 2009 Nov;76(5):1123–1129. doi: 10.1124/mol.109.060129

Fig. 3.

Fig. 3.

Rap1 is not coupled to ERK activation. A, WRT cells transfected with Rap1 siRNAs (Rap#1 and Rap#2) were starved, stimulated with forskolin for 2 min, and ERK phosphorylation, Rap1, and Akt expression (as a loading control) were analyzed. Rap1 expression was reduced by more than 80% after transfection with Rap1-directed siRNAs. B, the results from three experiments are summarized. C, starved cells were stimulated with EcAMP or TSH for the indicated times (in minutes). Lysates were subjected to RalGDS-RBD pull-down assay and blotted for Rap1GTP. Total cell lysates were analyzed for Rap1 expression. D, starved cells were stimulated with EcAMP for the indicated times (in minutes) and analyzed for ERK phosphorylation. Western blotting for actin was used to ensure equal protein loading.