Figure 1
Models for cold exposure in mice. (A,F) Acute mild cold exposure (A) and chronic (F) intermittent mild cold exposure protocols. Mice were exposed to 12 °C for 6 h in one day (left, acute protocol) or daily for 14 days (right, chronic protocol) and then sacrificed. Bottom rows of both protocols indicate controls. (B,G) Ucp1 mRNA expression in iWAT from control and cold exposure groups, normalized by 34B4 mRNA. n = 3–7. (C,H) Representative images of iWAT. Haematoxylin and eosin staining and Ucp1 immunostaining from control and cold exposure groups (left and right columns, respectively). (D,I) Immunoblot analysis of the indicated mitochondrial proteins in iWAT from control and cold exposure groups. HSP70 was used as a loading control. Data from two representative mice in each group are shown. (E,J) Densitometric analysis of mitochondrial proteins in iWAT from acute (D) and chronic (I) cold exposure groups. Levels were normalized to HSP70 as an internal control. (K) Quantification of miR-494-3p expression in iWAT from control and cold exposure groups and BAT from control groups, normalized by U6. n = 3–5. (L) Quantification of miR-494-3p and miR-494-5p expression in iWAT from control groups, normalized by U6. n = 8. *p < 0.05, ***p < 0.001. Full-length blots are presented in Supplementary Fig. 5. iWAT: inguinal white adipose tissue; Ucp1: uncoupling protein 1; PGC1-α: PPAR gamma coactivator 1-α; TFAM: transcription factor A mitochondria; PDH: pyruvate dehydrogenase E1-alpha subunit; ANT1/2: adenine nucleotide translocase 1/2; MTCO1: cytochrome c oxidase subunit 1; HSP70: heat shock protein 70; BAT: brown adipose tissue.
