Fig. 4: IR-61 stimulates mitochondrial function through ROS-Akt-Acly signaling.

a Immunoblots of Acly, p-Acly, Akt, and p-Akt in whole-cell lysates from the IR-61-treated BMDMs for the indicated times. β-Actin was used as the loading control. b Mitochondrial superoxide variation at 0–9 h after IR-61 treatment (n = 3). c Intracellular ROS variation at 0–9 h after IR-61 treatment (n = 3). d Immunoblots of Acly, p-Acly, Akt, and p-Akt in whole-cell lysates from the BMDMs treated with the vehicle control, NAC (2.5, 5, and 10 mM), or IR-61 with or without NAC for 6 h. β-actin was used as the loading control. e Immunoblots of respiratory chain subunits in whole-cell lysates from the IR-61-treated BMDMs for 72 h after transfection with siRNA control (siCtrl) or Acly siRNA1 (siAcly1). Gadph was used as the loading control. f Complex I, II, and IV activity was measured by ELISA of the BMDMs treated with IR-61 for 72 h after transfection with siCtrl or siAcly. g, h OCR and mitochondrial function of the BMDMs treated with siCtrl or siAcly for 48 h and then treated with IR-61 or vehicle control for another 24 h. i Pro-inflammatory mRNA levels of the IR-61-treated BMDMs transfected with siCtrl or siAcly for 48 h, and then treated with LPS for another 24 h. Data are representative of three independent experiments. Results are presented as the mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001, n = 3; two-sided Student’s t-test). Exact p-values are given in the Source Data file. Source data are provided as a Source Data file.