Fig. 1. Exposure of astrocytes to rTat/cocaine augment mitochondrial respiration.

a Extracellular lactate levels in media from astrocytes exposed to rTat, cocaine, or rTat and cocaine. b Representative Western blot of monocarboxylate transporter 4 (MCT4) levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. c Quantification of normalized intensity of MCT4 levels from (b) to GAPDH. d Quantification of LDH activity in astrocytes exposed to rTat, cocaine, or rTat and cocaine. e Quantification of intracellular lactate levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. f Quantification of pyruvate levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. g Quantification of PDH activity in astrocytes exposed to rTat, cocaine, or rTat and cocaine. h Representative western blot of PDHA1 (phospho S293) and PDH from astrocytes exposed to rTat, cocaine, or rTat and cocaine. i Quantification of normalized PDHA1 (phospho S293) levels to total PDH. j Quantification of Acetyl-CoA levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. k Measurement of oxygen consumption rate (OCR) in astrocytes exposed to rTat, cocaine, or rTat and cocaine. After basal OCR measurement, oligomycin (A), FCCP (B), and rotenone + Antimycin A (C) were added as indicated. Representative traces of OCR in astrocytes are shown. l, m Quantification of basal OCR (l), maximal OCR (m) in astrocytes exposed to rTat, cocaine, or rTat and cocaine. n Quantification of cellular ATP levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. o Representative Western blot for electron transport chain complex components (CI, CII, CIII, CVI, CV) in astrocytes exposed to rTat, cocaine, or rTat and cocaine. The outer mitochondrial membrane receptor, TOM20 was used as the loading control. p Quantification of mitochondrial ROS levels in astrocytes exposed to rTat, cocaine, or rTat and cocaine. Data indicate Mean ± SEM; ***P < 0.001, ** P < 0.01, *P < 0.05; n = 12–16