Fig. 12.

The relative hepatic content of eIF2α and eIF2αP and qRT-PCR analyses of Grp78 and Grp 94 mRNA content in untreated and PB-treated C57BL/6J WT and HRI(−/−) hepatocytes. A, lysates from H-depleted or H-repleted hepatocytes were assayed exactly as detailed under Materials and Methods. A prototype immunoblot is shown. Values from C57BL/6J WT are mean ± S.D. of three separate experiments or average of two separate experiments in the case of the C57BL/6J HRI (−/−). The interindividual variability between the two average values of two separate experiments each marked by the same symbol was >10%. B, hepatocytes were treated exactly as detailed under Materials and Methods. Lysates from H-depleted or H-repleted hepatocytes were assayed exactly as detailed in Fig. 8B. A prototype immunoblot is shown. The interindividual variability between the two average values of two separate experiments each marked by the same symbol was >10%. C, qRT-PCR analyses of Grp78 mRNA after heme depletion or repletion of untreated or PB-treated C57BL/6J HRI (−/−; KO) hepatocytes and corresponding HRI (+/+; WT) controls. Corresponding analyses of Grp78 mRNA from hepatocytes treated in parallel with thapsigargin (Tg), an established ER-stress inducer, are also included. Note the Y-axes scale differences. For experimental details, see Materials and Methods. Statistically significant differences were observed between the two mean ± S.D. values from three separate experiments each marked with the same symbol as follows: #, p < 0.05; §, p < 0.001; ‡, p < 0.05; ¶, p < 0.001; €, p < 0.05; =, p < 0.001; ●, p < 0.001; †, p < 0.001; *, p < 0.05; and **, p < 0.001. D, corresponding qRT-PCR analyses of Grp94 mRNA. Statistically significant differences between the two mean ± S.D. values from three separate experiments each marked with the same symbol were as follows: #, p < 0.05; §, p < 0.05; ‡, p < 0.001; ¶, p < 0.001; =, p < 0.05; ●, p < 0.05; †, p < 0.05; and *, p < 0.05.