Fig 2: PAQR11 regulated fasting-induced lipolysis in eWAT and lipid accumulation in liver. (A) The mRNA levels of Paqr11 and lipolytic genes in eWAT of wild-type mice at 12 weeks old fed with normal chow under fed and fasted (for 24 h) conditions as determined by quantitative RT-PCR. (B) Immunoblotting analyses of lipolytic proteins using eWAT from wild type (wt) and Paqr11−/− mice at 12 weeks old fed with normal chow under fed and fasted (for 24 h) conditions. (C) Liver triglyceride levels of the mice as in B. (D) Representative images of Oil Red O staining of the liver sections of the mice as in B. (E) The mRNA level of Paqr11 in eWAT of saline–or insulin-injected wild-type mice. Insulin was used at 0.6 U/kg. (F) Serum level of non-esterized fatty acid (NEFA) of the saline–or insulin-injected 12-week-old wt and Paqr11−/− mice fed with normal chow. (G) Immunoblotting to analyze phosphorylated HSL and total HSL of the mice as in F. (H) Immunoblotting to detect phosphorylated PKA substrates of the mice as in F. (I) A diagram to depict the working model of PAQR11 regulation on fasting-induced lipolysis in adipose tissue. All the quantitative data are shown as mean ± SD. ∗ for P < 0.05, ∗∗ for P < 0.01, ∗∗∗ for P < 0.001 and ns for non-significant.

Fig 3: Alterations in subcellular PRIP distribution and HSL phosphorylation in explant adipose tissues.(A, B) Adrenaline stimulation (Adrn) induces HSL translocation from cytosol fraction (sup) to fat-cake fraction (fat) in WT and PRIP-DKO explant adipose tissues. The bars for fat (black) and sup (white) in B represent the floating fat-cake and supernatant fractions, respectively. The amount of total HSL in fat and sup fractions is expressed as 100%. All the data represent mean ±SEM. **p<0.01 and n.s. (not significant) versus the corresponding WT value. Altered phosphorylation of HSL at Ser563 and Ser660 and perilipin at Ser492 in adipose explant lipolysis assays with or without stimulation by 1 µM adrenaline are also shown in (A). A representative image is shown; similar images were obtained from three independent experiments. (C, D) PRIP translocation to the lipid droplet fraction in response to 1 µM adrenaline (Adrn) stimulation. The bars for fat (black) and sup (white) in (D) represent the floating fat-cake and supernatant fractions, respectively. The amount of total PRIP in fat and sup fractions is expressed as 100%. Perilipin and β-actin are lipid droplet and cytosol marker proteins, respectively. A representative image is shown; similar images were obtained from three independent experiments (C). The data represent mean ±SEM. **P<0.01 and ***P<0.001 versus the corresponding values of PRIP1 and PRIP2 without adrenaline stimulation.

Fig 4: Effect of water-insoluble β-glucan on the expression of lipid metabolism-associated proteins in the liver tissues of HFD-fed mice. (a) Representative western blot for FAS and HSL. Quantification of FAS (b) and HSL (c) protein expression levels. NC, normal control group; HFD, obesity model group; Insoluble, mice fed HFD and given oral administration of water-insoluble β-glucan. Data are presented as mean ± standard deviation (n = 6). Groups labeled with different letters are different from one another (p < 0.05).

Fig 5: The in vivo effects of CL and CL + ghrelin co-injections on the activation of lipolytic enzymes ATGL and HSL in subcutaneous inguinal white (a) and visceral retroperitoneal (b) adipose tissue. Data were analyzed using a repeated measures one-way ANOVA (n = 6–8) and expressed as mean ± standard error, in arbitrary protein units (phospho/total). Data sharing a letter are not statistically different from each other. p < 0.05 was considered statistically significant.