Skip to main content
. 2010 Apr;77(4):575–592. doi: 10.1124/mol.109.061259

Fig. 2.

Fig. 2.

Effects of acute hepatic heme depletion and repletion on hepatic HRI autophosphorylation, eIF2α kinase activation, and PB-mediated CYP2B induction in cultured C57BL/6J mouse hepatocytes. Mouse hepatocyte cultures were untreated, heme (H)-depleted, or heme-repleted after heme depletion as detailed under Materials and Methods. A, a representative example of HRI Western immunoblotting analyses of these hepatocyte lysates (100 μg of protein) is shown at the top, with corresponding aliquots used for actin immunoblotting analyses as loading controls. Densitometric quantification of total hepatic HRI and autophosphorylated HRI (HRI-P) content and corresponding statistically significant differences between mean ± S.D. of three individual experiments are shown at the bottom. B, Western immunoblotting analyses of total eIF2α and eIF2αP content in these hepatocyte lysates (10 μg of protein) is shown at the top, with corresponding aliquots used for actin immunoblotting analyses as loading controls. Densitometric quantification of total hepatic eIF2α and eIF2αP content, and corresponding statistically significant differences between mean ± S.D. of 4 individual experiments are shown at the bottom. C, mouse hepatocyte cultures were untreated (first two lanes) or heme-depleted (H; next three lanes), pretreated with PB (next 12 lanes): alone (PB), with heme (H-control), heme depletion (H-depleted), or heme repletion after heme depletion (H-repleted), as detailed under Materials and Methods. CYP2B Western immunoblotting analyses of these hepatocyte lysates (30 μg of protein) is shown at the top, with corresponding aliquots used for actin immunoblotting analyses as loading controls. Densitometric quantification of hepatic CYP2B content from three individual experiments is shown at the bottom. Statistical analyses revealed significant differences in hepatic CYP2B content between untreated and PB-pretreated at p < 0.001, PB/H-depleted, and PB-pretreated at p < 0.001, PB/H-repleted, and PB/H-depleted at p < 0.001. No statistically significant differences were observed between PB-pretreated and PB/H-repleted. D, mouse hepatocyte cultures were treated as in C. TDO Western immunoblotting analyses of these hepatocyte lysates (20 μg of protein) is shown at the top, with corresponding aliquots used for actin immunoblotting analyses as loading controls. Densitometric quantification of hepatic TDO content as the average values from 2 individual experiments is shown at the bottom.