Fig. 1: miR-33^−/− mice exhibit impaired thermogenesis and BAT activation in a cold environment.

a Serial core body temperature change of miR-33^+/+ and miR-33^−/− mice kept at 4 °C. n = 10 and 11 mice per group, ***p < 0.001, two-way repeated-measures ANOVA with Bonferroni’s post hoc test. b Representative images of HE staining for BAT in miR-33^+/+ and miR-33^−/− mice kept at room temperature (RT). n = 6 mice per group. Scale bar, 1 mm (upper), 200 μm (lower). c Representative images of HE staining for BAT in miR-33^+/+ and miR-33^−/− mice kept at 4 °C for 6 h. n = 5 mice per group. Scale bar, 1 mm (upper), 200 μm (lower). d Quantitative PCR analysis of thermogenic genes in BAT of miR-33^+/+ and miR-33^−/− mice kept at room temperature (RT) and 4 °C for 6 h (cold). n = 6 mice per group at room temperature, n = 8–9 mice per group at 4 °C for 6 h. ***p < 0.001, **p < 0.01, *p < 0.05, two-sided Mann–Whitney test. e Representative images of HE staining for BAT of miR-33^+/+ and miR-33^−/− mice kept at 4 °C for 48 h. n = 6 mice per group. Scale bar, 200 μm. f Western blotting analysis of ABCA1, UCP1, TH, and β-actin in the BAT of miR-33^+/+ and miR-33^−/− mice kept at room temperature (RT) and 4 °C for 48 h (cold). n = 2, 3 mice per group. The arrow indicates the specific bands for TH staining. g Densitometry for UCP1 and TH in the BAT of miR-33^+/+ and miR-33^−/− mice kept at 4 °C for 48 h. n = 4 mice per group, *p < 0.05, two-sided Mann–Whitney test. h Representative images of immunohistochemistry for TH in the BAT of miR-33^+/+ and miR-33^−/− mice at room temperature. n = 3 mice per group. Scale bar, 100 μm. All data are presented as mean ± SEM.