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. 2020 Dec 4;21(12):e49634. doi: 10.15252/embr.201949634

Figure EV4. The malate‐aspartate shuttle is required for increased Norepinephrine‐stimulated energy expenditure induced by MPC inhibition.

Figure EV4

  1. Brown adipocytes were pre‐treated with vehicle (DMSO), 50 nM UK5099, aminooxyacetic acid (AOA) at various concentrations, or a combination. Oxygen consumption rates (OCR) were measured in respirometry media supplemented with 3 mM glucose and 3 mM glutamine in the presence of the compounds. Data show non‐stimulated and norepinephrine‐stimulated OCR (n = 3 individual experiments). Note that AOA has no effect in vehicle‐treated cells, but reduced OCR when cells are treated with UK5099.
  2. mRNA levels of OGC1 in brown adipocytes transduced with adenovirus carrying either scramble RNA (Scramble) or OGC1 shRNA (OGC1 KD). Data were normalized to vehicle for each individual experiment (n = 3 individual experiments). ****P < 0.0001 compared to Scramble by Student’s t‐test.
  3. Quantification of norepinephrine‐stimulated OCR after vehicle or UK5099 treatment in scramble RNA or OGC1 shRNA transduced cells. Data were normalized to vehicle for each individual experiment (n = 5 individual experiments). *P < 0.05 compared to vehicle by Student’s t‐test.
  4. mRNA levels of Aralar1 in brown adipocytes transfected scramble RNA (Scramble) or Aralar1 siRNA (Aralar1 KD). Data were normalized to vehicle for each individual experiment (n = 4 individual experiments). ***P < 0.001 compared to Scramble by Student’s t‐test.
  5. Quantification of NE‐stimulated OCR after vehicle or UK5099 treatment in scramble RNA or Aralar1 siRNA‐transfected cells. Data were normalized to vehicle for each individual experiment (n = 4 individual experiments). P = 0.0504 compared to vehicle by Student’s t‐test.

Data information: All data are presented as mean ± SEM.