FIGURE 9.

Subcellular fractionation of DGKη-PH in COS-7 cells treated and untreated with hyperosmotic shock. COS-7 cells expressing AcGFP alone and AcGFP-tagged DGKη-PH were lysed. The mixture was centrifuged at 12,000 × g for 5 min at 4 °C and further centrifuged at 100,000 × g for 60 min to separate supernatant (sup) and precipitate (ppt) fractions. A, AcGFP alone and AcGFP-tagged DGKη-PH were detected by Western blotting with anti-GFP antibody. B, the blots were scanned and quantified using ImageJ software. Relative intensities of precipitate and supernatant fractions of AcGFP alone and AcGFP-tagged DGKη-PH are indicated, respectively. Representative data of three independent experiments are shown.