Fig. 5: Cell autonomous regulation of energy expenditure by p62.
From: The scaffold protein p62 regulates adaptive thermogenesis through ATF2 nuclear target activation
Oxygen consumption rate (a), as well as basal and proton leak respiration (b) in BAT primary cells harvested from 6- to 8-wk-old male wt or p62Δ69-251 mice. Western blot analysis of p-ATF2 (Thr69/71) and UCP1 (c) and gene expression of Ucp1 and Pgc-1α (d) in 6 d differentiated BAT primary cells harvested from 6- to 8-wk-old male wt or p62Δ69-251 mice and which have been stimulated with saline (vhcl) or isoproterenol (0.5 μM) for 6 h. For measurement of oxygen consumption (a), basal respiration (b) and proton leak respiration (c), BAT primary cells were harvested from p62 wt or p62Δ69-251 mice, plated with equal number of cells in n = 4/5 wells each genotype and individually differentiated into mature brown adipocytes. Oxygen consumption was then measured in n = 4/5 individually differentiated wells each genotype with n = 3 technical replicates each well. For data in panel (c), BAT primary cells were harvested from p62 wt or p62Δ69-251 mice, plated with equal number of cells in n = 3 wells each treatment group and individually differentiated into mature brown adipocytes. Protein level of p-ATF2, GAPDH and UCP1 was then measured in n = 3/3/3/3 individually differentiated wells. For data in panel (d), BAT primary cells were harvested from p62 wt or p62Δ69-251 mice, plated with equal number of cells in n = 3 wells each treatment group and individually differentiated into mature brown adipocytes. Target gene expression was then measured in n = 3/3/3/3 individually differentiated wells with n = 2 technical replicates each well. Panels (a) and (d) are representative examples of two independently performed studies, each yielding similar results. Data in panel (a) have been analyzed using two-way ANOVA with time and genotype as co-variant and using Bonferroni post hoc multiple comparison to test for differences in individual time points. Bar graphs have been analyzed using two-sided two-tailed t-test. Data represent mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. Exact p-values are given in the Source data file.
