SoNar indicates metabolically distinct populations of FL hematopoietic cells. (A) The ratios of SoNar fluorescence (F405/F488 nm) were determined in CD45⁺SoNar⁺ FL hematopoietic cells by confocal microscopy, and representative images are shown. (B) Quantification of the ratios of SoNar fluorescence in panel A (n = 184). SoNar-low (ratio <0.6), SoNar-mid (ratio 0.6-1.8), and SoNar-high cells (ratio >1.8) were defined accordingly. (C) Representative images of the ratios of SoNar fluorescence with excitation at 405 and 488 nm (F405/F488 nm) in CD45⁺SoNar⁺ FL hematopoietic cells at indicated time points upon PBS, pyruvate, oxamate, AOA, or rotenone incubation. (D) Quantification of the ratios of SoNar fluorescence in panel C. A total of 25-55 CD45⁺SoNar⁺ FL hematopoietic cells were analyzed (n = 3). (E) Representative images of the ratios of SoNar fluorescence in CD45⁺ SoNar FL hematopoietic cells (E12.5) at indicated time points upon sequential treatments with PBS, pyruvate, and oxamate. (F) Quantification of the ratios of SoNar fluorescence in panel E. A total of 22 SoNar FL hematopoietic cells were analyzed (n = 3). (G) Representative flow cytometric analyses of the changes of the ratios of SoNar fluorescence in CD45⁺SoNar⁺ FL hematopoietic cells upon PBS, pyruvate, oxamate, AOA, or rotenone treatment. (H) Quantification of the ratios of SoNar fluorescence in panel G (n = 3). Scale bar, 10 μm. Data are represented as mean ± standard error of the mean. One-way analysis of variance with Tukey’s multiple comparison test was used for the comparison of statistical significance. See also supplemental Figure 2. ***P < .001.