Fig. 1: Expression of mitoNEET mRNA and protein increases during microbial sepsis.
From: Inhibition of mitoNEET attenuates LPS-induced inflammation and oxidative stress
Total RNA and protein were extracted from the spleen 48 h after sham or CLP surgery, and fibrin clot-induced microbial sepsis was triggered by E. coli or S. aureus bacteria (1 × 108 CFU). Expression of mitoNEET mRNA (A, C) and protein (B, D) levels was assessed by quantitative real-time RT-PCR or western blotting. *P < 0.05 for sham vs. CLP or fibrin clot-induced microbial sepsis. C57BL/6 mice were injected with LPS (20 mg/kg) or vehicle, and total RNA and protein were extracted from the spleen 6, 12, 24, 48, and 72 h after administration of vehicle or LPS (100 ng/mL). Total protein was extracted from BMDMs 3, 6, 12, and 24 h after administration of vehicle or LPS (100 ng/mL). Expression of mitoNEET mRNA (E) and protein (F, G) was assessed by quantitative real-time RT-PCR or western blotting. Total protein was extracted from BMDMs 6 h after administration of vehicle or LPS (100 ng/mL) plus a signaling inhibitor (5 µM BAY11-7082, 10 µM SP600125, 10 µM SB203580, 10 µM U0126, 10 µM LY2940002, or 20 mM NAC). Expression of mitoNEET protein was assessed by western blotting (H). All data are expressed as the mean ± SD from three independent experiments. *P < 0.05 for vehicle vs. LPS treatment.
