Fig. 6
From: Tip60-mediated lipin 1 acetylation and ER translocation determine triacylglycerol synthesis rate
Lipin 1 acetylation senses fatty acid abundance by its reciprocal association with Sirt1 and Tip60. a Lipin 1 dephosphorylation promotes its acetylation by Tip60. Flag-tagged WT-lipin 1 or 2KR-lipin1 or 16SA-lipin 1 was co-expressed with or without Tip60 in HEK293 cells, immunoprecipitated by antibody to Flag, followed by immunoblotting. b Dephosphorylation of lipin 1 increases its acetylation by Tip60. HEK293 cells transfected with vector, WT-lipin 1 or 2KR-lipin 1 or 16SA-lipin 1 (phosphorylation-defective) were treated with or without OA for 3 h, immunoprecipitated with antibody to Flag and immunoblotted as indicated. c Dephosphorylation of lipin 1 attenuates its association with Sirt1. HEK293T cells were transfected with Flag-lipin 1 and Myc-Sirt1, immunoprecipitated with antibody to Flag and immunoblotted as indicated. d Tip60 and Sirt1 compete with each other in the interaction with lipin 1. HEK293T cells were transfected with Flag-lipin 1, HA-Tip60 and varied amounts of Myc-Sirt1, immunoprecipitated with antibody to Flag and immunoblotted as indicated. e Interaction of endogenous Tip60 and Sirt1 with lipin 1. Lysates of 3T3-L1 adipocytes treated with or without OA for 3 h were immunoprecipitated by antibody to lipin 1 or Tip60 and analyzed by immunoblotting. f TAG/DAG production rates of WT or Tip60SA/SA adipocytes expressing shRNA targeting Lpin1 with reintroduction of WT-lipin 1, acetylation-defective (2KR) or acetylated mimetic (2KQ) mutant of lipin 1 (n = 4 experiments). Error bars denote SEM. Statistical analysis was performed by ANOVA followed by Tukey in f. *P < 0.05; N.S. not significant. Uncropped blots can be found in Supplementary Fig. 6
