Fig. 2.

On-plate subcellular fractionation of adherent human cells. A, Extraction procedure schematic for collection of cytoplasmic proteins (blue circles), nuclear proteins (orange and small blue circles), and transmembrane proteins (orange rectangles) from adherent cell cultures. HK, hexokinase. Gluc, glucose. G6P, glucose-6-phosphate. B–C, Immunoblot comparison of saponin extracts (SE), Nonidet P-40 extracts (NE) and insoluble proteins (I) for cytoplasmic (MEK1/2 and IκBα) and nuclear (PARP and JunD) proteins (B) as well as endogenous MKP3 phosphatase, the catalytic subunit of PP1 (PP1c) phosphatase, and the catalytic subunit of PP2A (PP2Ac) phosphatase (C) in the indicated human cell lines.