Mislocalization of RAS proteins in Icmt-deficient splenocytes and analyses of downstream RAS effectors in CD11b-positive splenocytes. (A) Intracellular localization of RAS proteins in splenocytes. Total cell extracts (T) from splenocytes were fractionated into soluble (S) and membrane (P) fractions and analyzed on Western blots with a pan-RAS antibody. Note the accumulation of RAS proteins in the soluble (S) fraction in splenocytes of Icmtfl/flKLSLM mice. (B) Western blots showing levels of GTP-bound RAS and downstream effectors in serum-starved and GM-CSF-stimulated CD11b-positive splenocytes pooled from mice of the same genotype (control, n = 5; Icmtfl/+KLSLM, n = 3; Icmtfl/flKLSLM, n = 3).