Fig. 3: miR-33^−/− mice exhibit reduced sympathetic nerve activity.

a Quantitative PCR analysis of thermogenic genes in denervated or sham-operated BAT kept at 4 °C for 5 h. Left side-BAT was denervated and right side-BAT was sham-operated in miR-33^+/+ and miR-33^−/− mice. At 1 week after the operation, mice were transferred to 4 °C for 5 h. n = 5 mice per group, ***p < 0.001, **p < 0.01, *p < 0.05, one-way ANOVA with Bonferroni’s post hoc test. b Norepinephrine content in BAT of miR-33^+/+ and miR-33^−/− mice before and after AMPT injection. n = 5–6 mice per group, *p < 0.05, two-way ANOVA with Bonferroni’s post hoc test. c Representative images of c-fos immunohistochemistry in the rRPa of miR-33^+/+ and miR-33^−/− mice kept at 4 °C for 2 h. n = 9 mice per group. Scale bar, 200 μm (upper). d Number of c-fos-positive cells in the rRPa of miR-33^+/+ and miR-33^−/− mice kept at 4 °C for 2 h. n = 9 mice per group, *p < 0.05, two-sided unpaired t test. e Quantitative real-time PCR analysis of beige adipocyte markers in inguinal WAT kept at 16 °C for 1 week in miR-33^+/+ and miR-33^−/− mice. n = 8 mice per group, **p < 0.01, *p < 0.05, two-sided unpaired t test. f Western blotting analysis of PGC1α, UCP1, TH, and β-actin in inguinal WAT at 16 °C for 1 week in miR-33^+/+ and miR-33^−/− mice. n = 4 mice per group. n = 4 mice per group. The arrow indicates specific bands of TH staining. g Densitometry for UCP1 and TH in the inguinal WAT of miR-33^+/+ and miR-33^−/− mice kept at 16 °C for 1 week. n = 4 mice per group, *p < 0.05, two-sided Mann–Whitney test. All data are presented as mean ± SEM.