Figure 1

PLD activity is decreased during 3T3-L1 adipogenic differentiation.

(a) 3T3-L1 cells were induced to differentiate for eight days. On day 0, 2, 4, 6 or 8 of differentiation, cells were lysed and a PLD assay was performed using a PLD assay kit. (b,c) (b) Recombinant PLD1 or PLD2 was transiently expressed. (c) Cells were differentiated for four days and recombinant in vivo PLD activities were measured using a transphosphatidylation assay as described in Materials and Methods. (d) Cells were treated as described in (a) and the lysates were subjected to western blotting. (e) White adipose tissue (WAT) was isolated from C57BL/6 J mice fed either a normal diet (ND) or a high-fat diet (HFD) for 12 weeks, lysed and subjected to PLD assay using a PLD assay kit (n = 4 per group). (f) WAT was isolated from either C57BL/6 J mice or ob/ob mice at age of 12 weeks and then subjected to PLD assay using a PLD assay kit (n = 9–10 per group). (g) Cells were differentiated in media under various conditions for four days and then in vivo PLD activities were measured with a transphosphatidylation assay. (h) 3T3-L1 cells were treated with 100 μM of 8-Br-cAMP and in vivo PLD activity was measured as in (g). (a–h) Data are representative of three to four independent experiments. Data are mean ± SD, with paired t-tests performed as indicated, *P < 0.05, **P < 0.01.