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. 2024 Apr 10;32(2):101249. doi: 10.1016/j.omtm.2024.101249

Figure 3.

Figure 3

MP TRAC-CAR T cells are less glycolytic and have increased mitochondrial mass

(A) ECAR and OCR for MP and Control TRAC-CAR T cells measured over time by Seahorse (oligomycin [2.5 μM], FCCP [1 μM], rotenone/antimycin A [0.5 μM]). (B) Basal/maximum respiration rates, spare respiratory capacity, and basal OCR/ECAR for MP and Control TRAC-CAR T cells measured by Seahorse. Two donors, NMP = 21, NControl = 6. (C) Dot plots for normalized mitochondrial mass (MitoTracker Green intensity divided by FSC-A), normalized mitochondrial membrane potential (TMRE dye intensity divided by FSC-A), cell size (FSC-A), and granularity (SSC-A) for CAR+ MP and Control TRAC-CAR T cells. Two donors, NMP = 4264, NControl = 2439. (D) Images and bar graphs of NAD(P)H mean lifetime (NAD(P)H τm) and free NAD(P)H fraction (NAD(P)H α1) of CAR+ MP and Control TRAC-CAR T cells as measured by fluorescence lifetime imaging. Two donors, NMP = 84, NControl = 37. OCR, oxygen consumption rate; ECAR, extracellular acidification rate; SSC-A, side scatter area; FSC-A, forward scatter area; TMRE, tetramethylrhodamine ethyl ester perchlorate. Error bars represent mean and standard deviation. Statistical significance was determined with unpaired t tests; ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.