Measurement for CCCP-induced thermogenesis in mouse brown adipocytes. (A) Left: Bright-field image of mouse brown adipocytes. Right: Fluorescence image of EuTTA for the cells on a RT nanosheet. (B) Time course of changes in the EuTTA/Rh101 ratio of RT nanosheets with mouse brown adipocytes upon application of 10 µM CCCP (red) or 0.1% DMSO (black). Values were corrected by those obtained in the medium (see Materials and methods for details). Number of cells, 59 (n = 6) and 31 (n = 3) for CCCP and DMSO, respectively. The horizontal black line indicates the period during which CCCP or DMSO was present. Calibration bar was calculated from the temperature sensitivity of RT nanosheets (−3.35%/°C, cf. Fig. 1 D). Data shown as mean ± SEM. (C) Left: Bright-field image of mouse brown adipocytes on a glass-bottom dish. Middle and right: Fluorescence images of ER thermo yellow and ER tracker green for the cells on left, respectively. (D) Time course of changes in the F.I. of ER thermo yellow in mouse brown adipocytes before and after application of 10 µM CCCP (red) or 0.1% DMSO (black). (E) Same as in D for ER tracker green. In D and E, number of cells, 27 (n = 3) and 22 (n = 2) for CCCP and DMSO, respectively. Data shown as mean ± SEM.