FIG. 2.

Pex5 knockdown exacerbates ROS and inflammation in 3T3-L1 adipocytes. (A–C) 3T3-L1 adipocytes were transfected with Pex5 siRNAs for 72 h. Reductions of Pex5 mRNA and protein expressions were verified. Gene and protein expressions related to peroxisome biogenesis and function were measured. Expression levels of indicated proteins were quantified by densitometry and normalized with GAPDH. (D) Immunostaining and (E) fluorescence-intensity quantification of Abcd3 in control and Pex5 knockdown adipocytes. Original magnification: 630 × ; scale bar: 10 μm. (F) Measurement of adipose tissue peroxisomal FAO and (G) OCR in the presence of 10 μM hexacosanoic acid (C26:0). (H) Cellular ROS was detected with DCF-DA and (I) staining intensity was quantified. Original magnification: 100 × ; scale bar: 100 μm. (J) Relative mRNA levels of inflammatory cytokines are shown. (K) Secreted IL-6 in cell culture supernatant was measured by enzyme-linked immunosorbent assay. (A–K) Values are mean ± SE of 3–4 experiments. *p < 0.05 versus control. DCF-DA, 2′,7′–dichlorofluorescein diacetate; DPM, disintegrations per minute; FAO, fatty acid oxidation; OCR, oxygen consumption rate; Pex, peroxisomal biogenesis factor; ROS, reactive oxygen species. Color images are available online.