Fig. 5.

Membrane association of cPLA₂γ is not determined by the C-terminal region or farnesylation. A: Cell homogenates of IMLF^−/− and SkMc cells infected with Ad-cPLA₂γ and incubated with or without BMS-214662 were prepared as described in Methods. The relative amount of cPLA₂γ in the 100,000 g soluble (cytosol; C) and particulate (membrane; M) fractions was determined by Western blot analysis of 30 μg of total protein per lane. B: Cell homogenates of COS cells overexpressing cPLA₂γ with the C-terminal CaaX sequence mutated were prepared as described in Methods. Amounts of mutated cPLA₂γ in the soluble (C) and particulate (M) fractions were determined by Western blotting. C: Cell homogenates of HEK cells overexpressing an ~55 kDa cPLA₂γ with the C terminus truncated were prepared as described in Methods. Amounts of mutated cPLA₂γ in the soluble (C) and particulate (M) fractions were determined by Western blotting.