Fig. 5: PEX5 escorts ATGL to LD to mediate fasting-induced lipolysis.

a Relative glycerol release from adipocytes transfected with negative control (NC) or PEX5 siRNA(siPEX5) for 48 h. n = 3 for each group. b Relative glycerol release from adipocytes transfected with siNC or siPEX5 for 48 h together in the absence or presence of WY-14643 (10 μM) treatment. n = 3 for each group. c Relative glycerol release from adipocytes transfected with siNC or siACOX1 for 48 h. n = 3 for each group. d, e Representative SIM images and quantification analysis of recruited ATGL to LDs in adipocytes immunostained with endogenous PLIN1 (red) and ATGL (green). Cells were transfected with siNC or siPEX5. n = 10 cells for siNC group; n = 15 cells treated with ISO; n = 12 cells for siPEX5 group; n = 13 cells for siPEX5 treated with ISO. Quantification of ATGL recruitment to LDs was measured using imageJ software. f Representative SIM z-section images (left) and fluorescence intensity profiles from the indicated line scans (right). Below 0.2 fluorescence intensity indicates background fluorescence signal. LD areas are highlighted in yellow. g Western blot of whole cell extracts (WCE) or LD fractionation from adipocytes transfected with siNC or siPEX5. 30 μg of protein from WCE; 20 μg of protein from LD fraction. h Quantification of ATGL in LDs normalized to PLIN1 from g. n = 4 independent experiments. CON control, ISO isoproterenol. Cells were treated with ISO (1 μM) for 1 h. All scale bars, 10 μm. Data represent the mean ± SD; ^*P < 0.05, ^***P < 0.01 in two-way ANOVA followed by Turkey’s post-hoc test. n.s., not statistically significant.