Fig. 6

AdipoR2 overexpression causes decreased dihydroceramide levels (data from the R2OE experiment). Western blots detection of the HA-tagged over-expressed AdipoR2 using an anti-HA antibody (A) or an anti-AdipoR2 antibody (B); anti-GAPDH was used as a loading control. (D-F) Relative abundance of SFA, MUFA and PUFA in phosphatidylcholines (PC) and phosphatidylethanolamines (PE) of control (WT) and AdipoR2 over-expressing cells (R2OE) treated with 400 μM PA for 6 h. The sums of five sphingolipid classes are shown as box plots (J-N) while their fatty acid composition is shown as a heat map (O). In the box plots, boxes indicate the 25th to 75th percentile while the whiskers indicate the data points still within 1.5 of the box range. For D-N, significant differences from the WT control were determined using Student’s t-tests, with *P < 0.05, **P < 0.01 and ***P < 0.001. For the heat map, the amount of each lipid species was normalized to the average of the WT + 400 μM PA treatment and the heat maps show fold differences from the mean across all treatments (each column is a replicate); only lipid species with significantly different levels among siRNAs in a given culture condition are included (ANOVA, q < 0.05)