Figure 4. Intracellular Ca 2 dynamics... | Rockefeller University Press

Figure 4.

Intracellular Ca2+dynamics regulates thermogenic gene program in white adipocytes. (A) Schematic showing the monitoring of Ca2+ activity in adipose tissue in vivo using the GCaMP6f transgenic mice. (B) Representative time-lapse images of iWAT of male GCaMP6f transgenic mice at different time intervals after 1 mg/kg NE stimulation. The corresponding video is included as Video 1. Scale bars: 50 μm. (C) Quantitative analysis of the GCaMP6f fluorescence intensity (Ft/F0) in B. Curve depicts the measurements from four independent mice. (D) Representative time-lapse images of differentiated white adipocytes isolated from iWAT of male GCaMP6f transgenic mice at different time intervals after DMSO (top) or 1 μM NE stimulation (bottom). The corresponding video is included as Video 2. Scale bars: 50 μm. (E) Quantitative analysis of the GCaMP6f fluorescence intensity (Ft/F0) in D. Curves depict the measurement of 100–141 cells from four independent experiments. (F) Gene expression (qRT-PCR) in differentiated white adipocytes. Cells were treated with 2 μM TG for 4 h. Inset: Quantitative analysis of the GCaMP6f fluorescence intensity (Ft/F0). n = 4 independent experiments. (G and H) Gene expression (qRT-PCR) in differentiated white adipocytes. Cells were treated for 4 h with DMSO (vehicle), 10 μM FSK, 20 μM CL, 10 μM BAPTA-AM, FSK + BAPTA-AM, or CL + BAPTA-AM as indicated in the key. n = 4 independent experiments. Values represent mean ± SEM. *, P < 0.05; **, P < 0.01; *** P < 0.001. P value was determined using two-tailed unpaired Student’s t-test (E and F) or one-way ANOVA coupled to a Fisher’s LSD post-hoc test (G and H). Data are representative of four independent experiments (B–H).

Intracellular Ca ²⁺ dynamics regulates thermogenic gene program in white adipocytes. (A) Schematic showing the monitoring of Ca²⁺ activity in adipose tissue in vivo using the GCaMP6f transgenic mice. (B) Representative time-lapse images of iWAT of male GCaMP6f transgenic mice at different time intervals after 1 mg/kg NE stimulation. The corresponding video is included as Video 1. Scale bars: 50 μm. (C) Quantitative analysis of the GCaMP6f fluorescence intensity (F_t/F₀) in B. Curve depicts the measurements from four independent mice. (D) Representative time-lapse images of differentiated white adipocytes isolated from iWAT of male GCaMP6f transgenic mice at different time intervals after DMSO (top) or 1 μM NE stimulation (bottom). The corresponding video is included as Video 2. Scale bars: 50 μm. (E) Quantitative analysis of the GCaMP6f fluorescence intensity (F_t/F₀) in D. Curves depict the measurement of 100–141 cells from four independent experiments. (F) Gene expression (qRT-PCR) in differentiated white adipocytes. Cells were treated with 2 μM TG for 4 h. Inset: Quantitative analysis of the GCaMP6f fluorescence intensity (F_t/F₀). n = 4 independent experiments. (G and H) Gene expression (qRT-PCR) in differentiated white adipocytes. Cells were treated for 4 h with DMSO (vehicle), 10 μM FSK, 20 μM CL, 10 μM BAPTA-AM, FSK + BAPTA-AM, or CL + BAPTA-AM as indicated in the key. n = 4 independent experiments. Values represent mean ± SEM. *, P < 0.05; **, P < 0.01; *** P < 0.001. P value was determined using two-tailed unpaired Student’s t-test (E and F) or one-way ANOVA coupled to a Fisher’s LSD post-hoc test (G and H). Data are representative of four independent experiments (B–H).