Extended Data Fig. 4: Mitochondrial properties of mouse and human PSM cells.
From: Metabolic regulation of species-specific developmental rates
a. Mass-specific oxygen consumption rate measured over the course of the mitochondrial stress test for MSGN1-Venus+ PSC-derived mouse (n = 9) and human (n = 7) PSM cells. 1 μM oligomycin, 1 μM FCCP, and 0.5 μM Rotenone + 0.5 μM Antimycin A were added at the timepoints marked by dotted lines. The first three timepoints denote basal respiration;, respiration after oligomycin addition corresponds to proton leak; FCCP induces maximal respiration; and rotenone/antimycin reveal non-mitochondrial respiration. Spare capacity refers to the difference between maximal and basal respiration rates. Mean ±SD. b. Spare respiratory capacity in MSGN1-Venus+ PSC-derived mouse and human PSM cells. Mean ±SD. n = 7 biological replicates. Unpaired two-sided t-test: p = 2.7 × 10−9. c. Oxygen consumption rate profiles for mitochondria isolated from mouse and human PSC-derived PSM cells. Rates correspond to 10 ug mitochondria seeded per assay well. ETC complex I substrates pyruvate and malate were provided to fuel respiration. 2 mM ADP, 5 μM Oligomycin, 6 μM FCCP, and 1 μM Rotenone with 1 μM Antimycin A were injected at the timepoints marked by dotted lines. First two timepoints correspond to basal respiration (state 2), followed by ADP-stimulated respiration (state 3), then leak respiration (state 4o), followed by maximal FCCP-stimulated respiration (state 3u), and finally non-mitochondrial respiration. Mean ±SD. n = 5 biological replicates. d. Whole-cell NAD+/NADH ratio in MSGN1-Venus+ PSC-derived mouse and human PSM cells. Each datapoint represents the average of 3 technical replicates. Mean ±SD. n = 6 biological replicates. Unpaired two-sided t-test: p = 1.2 × 10−7. e. Whole-cell ADP/ATP ratio in MSGN1-Venus+ PSC-derived mouse and human PSM cells. Mean ±SD. n = 9. Each datapoint represents the average of 3 technical replicates. Unpaired two-sided t-test: p = 2.42 × 10−7. f. Percent viable cells as measured by trypan blue staining in human PSM cells treated with the indicated inhibitors for 24 h. n = 3 independent experiments.
