Fig. 2

Tip60 plays an essential role in fatty acid-induced TAG synthesis. a Survival rates of Tip60^SA/SA litters nursed by WT or Tip60^SA/SA females (n = 8 litters for each group), and WT litters nursed by WT or Tip60^SA/SA females (n = 7 litters for each group). b Representative BODIPY staining (n = 3 experiments) of milk fat globules (green) from the mammary glands of early lactating (day 2 postpartum) WT and Tip60^SA/SA females. Scale bar, 50 μm. c Level of TAG in milk collected from WT and Tip60^SA/SA females (day 2 postpartum, n = 5 per group). d Representative images of mammary glands from WT and Tip60^SA/SA females at day 2 postpartum (n = 3 experiments), visualized by H&E staining. The WT mouse (left) has numerous fat globules in the apical regions of mammary epithelial cells and in the ductal lumens, but were rare in the Tip60^SA/SA mouse (right). Scale bars, 50 μm. e Lentivirus-mediated knockdown of Tip60 in 3T3-L1 adipocytes. TCL total cell lysate. f TAG and phosphatidylcholine (PC) synthesis rates of 3T3-L1 adipocytes expressing control shRNA (ctrl) or shRNAs targeting Tip60 with ³H-labeled oleic acid (OA) treatment (n = 4 experiments). g TAG and PC synthesis rates of 3T3-L1 adipocytes treated with DMSO (ctrl) or Tip60 inhibitor MG149 (n = 4 experiments). h Rates of diacylglycerol (DAG) and TAG synthesis from ³H-OA in eWAT explants isolated from 6-week-old WT and Tip60^SA/SA mice (n = 4 individuals per group). i Fatty acid oxidation rates of control (ctrl) or Tip60 knockdown or MG149-treated 3T3-L1 adipocytes or adipocytes derived from WT or Tip60^SA/SA MEFs (day 8) (n = 4 experiments). Error bars denote SEM. Statistical analysis was performed by two-tailed unpaired Student’s t-test in c, g, h and middle and right bar graph of i, by one-way ANONA followed by Tukey in f and left bar graph of i. *P < 0.05; **P < 0.01; N.S. not significant. Uncropped blots can be found in Supplementary Fig. 6