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. 2001 Feb;21(4):1173–1184. doi: 10.1128/MCB.21.4.1173-1184.2001

FIG. 5.

FIG. 5

Regulation of ERK phosphorylation by Rho family proteins. (A) Myocytes were transfected with ERKMyc and inhibitory mutants of Rho family proteins. Following exposure to ET-1 (100 nM, 5 min), ERKMyc was immunoprecipitated and immunoblotted with antibodies to phosphorylated ERK (upper panel). Parallel blots were stripped and probed with an antibody to the c-Myc tag (lower panel). The experiment was repeated on three separate occasions with similar results. (B) Myocytes were transfected with ERKMyc, ΔN-Raf, and/or activated mutants of Rho family proteins. When applicable, exposure to ET-1 (100 nM) was for 5 min. ERKMyc was immunoprecipitated and immunoblotted with antibodies to phosphorylated ERK (upper panel). Parallel blots were reprobed with an antibody to the c-Myc tag (lower panel). The experiment was repeated on four separate occasions with similar results. (C) Blots shown in panel B were analyzed by scanning densitometry, and the amount of phospho-ERKMyc was adjusted for total ERKMyc protein. Results are means ± SEM for four separate myocyte preparations. ∗, P < 0.05 relative to ΔN-Raf alone (unpaired two-tailed t test).