Research Paper Volume 9, Issue 10 pp 2069—2082

Figure 2. Celastrol differentially affected the expression of macrophage M1 and M2 biomarkers in epididymal adipose tissues and liver. (A) Detection of iNOS, COX-2, and arginase-1 in epididymal adipose tissues. After 21-day treatment, epididymal fat pads were recovered from mice, and stained with specific antibodies. CD68 was stained as pan-macrophage biomarker whereas the cell nuclei were stained with DAPI. The sections were imaged under a Zeiss LSM 780 confocal microscopy. Representative images were shown. Scale bar, 50 μm. (B) qRT-PCR quantification of macrophage M1 and M2 biomarkers in epididymal adipose tissues. Total RNAs were extracted from adipose tissues and analyzed by qRT-PCR technique using QuantiTect SYBR Green PCR Kit and specific DNA primers from Qiagen. N = 3; HFD, HFD only; C7.5, celastrol (7.5 mg/kg/d). (C) Detection of iNOS, COX-2, and arginase-1 in liver tissues. Livers were recovered from mice, and stained with specific antibodies. CD68 was stained as pan-macrophage biomarker whereas the cell nuclei were stained with DAPI. The sections were imaged under a Zeiss LSM 780 confocal microscopy. Representative images were shown. Scale bar, 50 μm. (D) qRT-PCR quantification of macrophage M1 and M2 biomarkers in liver tissues. Total RNAs were extracted from livers and analyzed by qRT-PCR technique using QuantiTect SYBR Green PCR Kit and specific DNA primers from Qiagen company. N = 3; HFD, HFD only; C5, celastrol (5 mg/kg/d); C7.5, celastrol (7.5 mg/kg/d); *, p < 0.05; **, p < 0.01; ***, p < 0.001.