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. Author manuscript; available in PMC: 2014 Jul 31.
Published in final edited form as: Sci Signal. 2013 Dec 3;6(304):ra105. doi: 10.1126/scisignal.2004125

Fig. 3. Effects of the knockdown of RasGRP isoforms on ERK phosphorylation in BMMPCs from WT and Mx1-Cre, KrasG12D mice.

Fig. 3

(A) Rasgrp3 and Rasgrp4 mRNA and RasGRP3 and RasGRP4 in BMMPCs from WT and KrasG12D mice were determined by quantitative RT-PCR. (B) Protein abundances were assessed by Western blotting (see also fig. S14) (C) BMMPCs that were incubated with or without siRNAs (10 nM) specific for Rasgrp3 (RG3) or RasGrp4 (RG4) or with a control scrambled siRNA (Sc) were stimulated with GM-CSF for 15 min. The cells were lysed and Western blotting analysis was performed to determine the amounts of RasGRP3, RasGRP4, and pERK, with actin used as a loading control. (D) BMMPCs were starved of serum and cytokines overnight, incubated with PI-90, U73122, KN-62, or PD0325901 for 30 min, and then stimulated for 15 min with GM-CSF alone or in the presence of 10 μM PLX4720. Cells were then lysed and analyzed by Western blotting to determine the abundances of pAkt and pERK, with actin used as a loading control. Data from additional independent experiments are presented in fig. S15.