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

Figure 1.

Figure 1

Chronic cold exposure increases autophagy in BAT and primary brown adipocytes

(A) Representative immunoblots and densitometry analysis of MAP1LC3BII, SQSTM1, and MAP1LC3BII:MAP1LC3BI ratio in BAT of mice subjected to 72 h of cold exposure.

(B) Immunoblots and densitometry analysis of PRKAA and MTOR pathway in BAT during cold exposure.

(C) mRNA expression of autophagy genes in the BAT of mice exposed to cold. Values are means ± SEM for 5 mice in each group.

(D) Autophagy flux analysis in primary brown adipocytes. Cells were treated with or without NE for 72 h, and bafilomycin (Baf) was added 6 h before harvest.

(E) Confocal microscopic images and quantification of red puncta in brown adipocyte cell line transfected with RFP-eGFP-MAP1LC3B plasmid and incubated 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.

(F) mRNA expression of autophagy genes in primary brown adipocytes treated with 1 μM NE for 72 h. Statistical significance shows as ∗p < 0.05.