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25 Prior to the pandemic, fentanyl mainly affected urban areas in the eastern regions of the U.S

All standards were obtained from Cambridge Isotope Laboratory unless stated otherwise

Cells were then washed twice with mitochondrial assay solution (220 mM mannitol [Sigma-Aldrich, M4125], 70 mM sucrose [Sigma-Aldrich, S7903], 10 mM KH 2 PO 4 [VWR, 470302], 2 mM HEPES [Fisher Scientific, BP310], and 1 mM EGTA [VWR, 97062]) and then overlaid with 175 L of mitochondrial assay solution supplemented with 10 mM sodium pyruvate (Sigma-Aldrich, P5280), 1 mM malate (Sigma-Aldrich, M0875), 4 mM ADP (Sigma-Aldrich, A5285), and Seahorse Plasma Membrane Permeabilizer (Agilent, 102504-100)