Fig. 6: Hypothermic defects were reproducible in EibKO mice. | Nature Communications

Fig. 6: Hypothermic defects were reproducible in EibKO mice.

From: Ei24 deficiency in brown adipocytes induces severe hypothermia under cold stress independent of UCP1 activity

Fig. 6: Hypothermic defects were reproducible in EibKO mice.The alternative text for this image may have been generated using AI.

a Western blot analysis of protein lysates obtained from BAT of male wild-type control and EibKO mice (representative results reproduced independently at least twice). b Images of interscapular BAT (left panel) isolated from male and female wild-type control and EibKO mice, and the calculated cross-sectional areas (right panel) based on the images shown in left panel (n = 6 biological replicates per group). c H & E staining (right panel) and LD size analysis (left panel) of iBAT and inguinal WAT from male wild-type and EibKO mice (n = 3 biological replicates per group; 2000 LDs for BAT, 90 for iWAT; Scale bars, 20 μm). d Core body temperatures of wild-type control and EibKO mice were measured at 25 °C and in a 4 °C environment. e Thermal images (left panel) and average surface temperature (right panel) of wild-type control and EibKO mice were measured at 25 °C and in a 4 °C environment. f Cellular ATP concentrations were measured in differentiated BAT SVFs from wild-type, EiaKO, and EiaKO-Ei24 re-expressed (RE) cells. g Mitochondrial matrix pH was measured in differentiated BAT SVFs from wild-type, EiaKO and EiaKO-RE mice (left panel). The resting mitochondrial matrix pH was quantified in differentiated BAT SVFs from wild-type, EiaKO, and EiaKO-RE mice (right panel). h Relative mitochondrial potentials (ΔΨm) were measured based on the ratio of tetramethylrhodamine methyl ester (TMRM) fluorescence to MitoTracker Green (MTG) fluorescence. Data in f, g are representative of 3 independent biological replicates. i Schematic summary of the effects of Ei24 deficiency in brown adipocytes. EiaKO BAT displayed enlarged lipid droplets and mitochondrial defects characterized by reduced membrane potential (ΔΨm), lowered mitochondrial matrix pH, and impaired fatty acid oxidation (FAO), leading to cold intolerance. Despite these defects, UCP1 expression and its activity remained comparable to controls. Created with Biorender.com56. The data shown represent the mean ± SEM. Statistical analysis was performed using an unpaired two-tailed Student’s t-test for the data shown in a-c, one-way ANOVA in d, g and h, and two-way ANOVA for the data shown in e, f.

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