Figure 6.

HILPDA alleviates ER stress in adipose tissue during refeeding. A) Relative Hilpda mRNA expression in epididymal white adipose tissue of Hilpda^flox/flox and Hilpda^ΔADIPO mice after 24 h of fasting or 20 h of fasting followed by 4 h of refeeding with chow (n = 9–13 per group). B) HILPDA, ATGL, and HSP90 protein levels in epididymal white adipose tissue. C) mRNA levels of Hilpda, Adipoq (adipocyte marker), and Cd14 (endothelial marker) in epididymal white adipose tissue, freshly separated adipocytes, and the stromal vascular fraction of Hilpda^flox/flox (n = 4) and Hilpda^ΔADIPO mice (n = 4). Gene expression levels of Hilpda in adipose tissue from Hilpda^flox/flox mice were set at one. D) Bodyweight of Hilpda^flox/flox and Hilpda^ΔADIPO mice after 24 h of fasting or 20 h of fasting followed by 4 h of refeeding with chow (n = 9–13 per group). gWAT, gonadal (epididymal) adipose tissue; iWAT, inguinal adipose tissue; BAT, brown adipose tissue. E) Weight of the liver and various adipose tissue depots. F) Plasma metabolites. G) Liver triglyceride content (%, wt/wt). H) Relative expression of ER stress genes in epididymal white adipose tissue. I) Protein levels of eIF2α, phosphorylated eIF2α, and BIP in epididymal white adipose tissue from fasted/refed Hilpda^flox/flox and Hilpda^ΔADIPO mice. Right lane is positive control of mouse adipocytes treated with thapsigargin. Western blots were probed with antibodies against eIF2α, p-eIF2α, BIP, HILPDA and HSP90. In the graphs, the horizontal bar represents the mean and the error bars represent SEM. P values in the figures reflect the statistical significance of the comparison Hilpda^flox/flox versus Hilpda^ΔADIPO by two-way ANOVA. Asterisk indicates significantly different from Hilpda^flox/flox mice according to Tukey's posthoc test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.