Skip to main content
. Author manuscript; available in PMC: 2019 Nov 6.
Published in final edited form as: Nat Metab. 2019 May 6;1(5):560–569. doi: 10.1038/s42255-019-0066-3

Fig. 6. PNPLA3 I148M variant is a gain of function for suppressing ABHD5-dependent lipolysis.

Fig. 6.

(a) Competitive BiFC assay in lipid-loaded COS-7 cells transfected with Yn-ABHD5 and PNPLA2-Yc along with either mCherry, WT PNPLA3-mCherry, PNPLA3 I148M-mCherry, or Plin1-mCherry. Data are biological quadruplicates from four independent experiments. *p=0.0397 and ***p=0.0001 indicates a difference compared to mCherry control, and &p<0.0312 indicates a significant difference between WT PNPLA3 and I148M as determined by one-way ANOVA with Tukey’s post t-test. Glycerol (b) and free fatty acid (FFA; c) release from Basal (no Dox, DMSO) or SR-3420 (20 μM, 1 hour) stimulated WT PNPLA3-mCherry or PNPLA3 I148M-mCherry brown adipocytes (BA) treated with differing doses of doxycycline (Dox; 0, 30, and 300 ng/ml). p values indicate a significant difference in glycerol release between WT PNPLA3 and PNPLA3 I148M as determined by two-way ANOVA with Bonferroni post t-test. &&&p=0.0001 indicates a main effect of doxycycline on FFA release as determined by two-way ANOVA. Data are biological quadruplicates from four independent experiments (d) Triacylglycerol (TAG) levels from control (-Dox) and doxycycline (+Dox) treated wildtype (WT) PNPLA3-mCherry and PNPLA3 I148M-mCherry BAs. (e) TAG levels from Control BA and BA cells with stable knockdown of ABHD5 (ABHD5 shRNA) expressing PNPLA3 I148M (+Dox). (d and e) Data are from one experiment with biological triplicates and are representative of three independent experiments. Data are expressed as means +/− SEM.