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. 2019 Sep 27;39(20):e00210-19. doi: 10.1128/MCB.00210-19

FIG 4.

FIG 4

In BAT, thermoneutral condition-induced LD unilocularization downregulates insulin-dependent GLUT4 translocation to the PM. Primary brown adipocytes were isolated from BATs of 8-week-old, male, C57BL/6J mice housed under thermoneutral (30°C), room temperature (25°C), or cold (4°C) conditions for 1 week. (A to C) F-actin (phalloidin-TRITC, red) and PM-anchored GLUT4 (green) of primary brown adipocytes were assessed by immunohistochemistry in the absence or presence of insulin (100 nM, 30 min). Scale bar, 50 μm (A). The average diameters (B) and intensities of PM-anchored GLUT4 (C) of isolated primary brown adipocytes were determined. (D) F-actin structure (phalloidin-TRITC, red) of BAT. Scale bars, 50 μm. Insets show ×4 magnifications of the indicated areas (nucleus [Hoechst, blue]). (E) F/G-actin ratios in primary brown adipocytes isolated from BAT. WCL, whole-cell lysates. Data represent means ± the SD. *, P < 0.05; ***, P < 0.001 (Student t test).