Fig. 1

4-OI decreases LPS-induced nephrotoxicity in vivo and apoptosis in vitro. Scr (A) and BUN (B) levels of different groups of mice exposed to LPS or 4-OI (n = 6). (C) qPCR analysis of mRNA of Lcn2  in renal tissues (n = 6). (D) Representative images of hematoxylin and eosin (H&E) staining of the renal cortex. Vacuolization and loss of brush border in tubular cells are pointed to by the arrows. Scale bar: 50 μm. (E) Tubular injury is evaluated by figuring out the renal tubules with signs of injury (n = 6). (F) Representative western blot bands of BAX, cleaved caspase-3, BCL2, and β-actin (protein loading control) in renal cortex tissues. (G) Relative band density of BAX, cleaved caspase-3 and BCL2 signals (n = 6). (H) In Hoechst 33258 staining images, nuclear condensation, fragmentation, and apoptotic bodies were observed. Scale bar = 100 μm. (I) The apoptotic index (AI) was calculated as the percentage of apoptotic nuclei per total nuclei number per field. Representative western blot bands (J) and relative band intensity (K) of BAX, cleaved caspase-3 and BCL2 in BUMPT cells treated with different doses of 4-OI under saline or LPS conditions (n = 4). Each symbol (circle) represents an independent experiment. Data are presented as means ± SDs. *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant. LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; Scr, serum creatinine; BUN, blood urea nitrogen; qPCR, Real-time polymerase chain reaction