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. 2021 Apr 15;24(5):102434. doi: 10.1016/j.isci.2021.102434

Figure 3.

Figure 3

Chronic cold exposure induces mitophagy and mitochondrial turnover in BAT

(A) Representative electron microscopic images showing BAT of mice housed at TN (30°C) or in cold (4°C) for 72 h. White box shows an autophagosome containing mitochondria. Scale bar, 0.2 μm.

(B) Immunoblots and densitometry of autophagy proteins in mitochondrial fraction of BAT.

(C) Confocal microscopic images and quantification of red puncta in brown adipocyte cell line transfected with mito-RFP-EGFP plasmid and treated with or without 1 μM NE for 72 h. Scale bar, 20 μm. Quantification of images (at least 10 transfected cells per sample in 3 different fields) was done using ImageJ software.

(D) Immunoblots and densitometry of mitochondrial proteins in BAT.

(E) Confocal microscopic images showing brown adipocyte cell line transfected with pMitoTimer plasmid and treated with or without 1 μM NE for 72 h. Bafilomycin (Baf) was added 6 h before harvest. Scale bar, 20 μm.

(F) Seahorse analysis of oxygen consumption rate (OCR) for primary brown adipocytes treated with 1 μM NE for 72 h. For the NE-treated group, 1 μM NE was added to the assay media 1 h before OCR measurement.

(G) Representative immunoblots and densitometry showing mitochondrial protein expression in primary brown adipocytes treated with 1 μM NE for 72 h. Statistical significance shows as ∗p < 0.05, ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001.