Figure 2

PLD1 negatively regulates adipogenic differentiation.

(a) 3T3-L1 cells were infected with lentiviruses expressing two different PLD1 shRNAs or scrambled shRNA, and then selected by puromycin for 5 days. Cells were lysed and subjected to western blotting. Western blot intensities were analyzed using ImageJ. (b,c) Cells were treated as described in (a) and induced into differentiation for eight days. Cells were either (b) stained with Oil Red O and quantified for lipid contents, or (c) lysed and subjected to western blotting. (d,e) Pld1^+/+ and Pld1^−/− MEFs were differentiated for eight days and either (d) subjected to staining with Oil Red O and quantified for lipid contents, or (e) lysed and analyzed by western blotting. (f,g) Cells were induced to differentiate three days after the infection of adGFP or adPLD1, followed by either (f) Oil Red O staining and quantification for lipid content, or (g) lysed and analyzed by western blotting. (a–g) Images are representative of three to five experiments. Scale bars, 100 μm. All data shown are mean ± SD or blots representative of three to five independent experiments. Student t-tests were performed to compare the indicated pairs of data. *P < 0.05; **P < 0.01.