Fig. 9.

qRT-PCR validation of polysome distribution of selected mRNAs in Perk wild-type vs. knockout perfused livers. Mouse livers were perfused for 35 min ± tBuHQ. Total RNA was extracted from the unfractionated supernatant and the nonpolysomal, subpolysomal, and polysomal gradient fractions for use in hybridization array and qRT-PCR experiments. Expression of mRNA was determined by qRT-PCR and normalized to the average expression of the housekeeping genes β-actin, Gapdh, and Tbp (see materials and methods for details on calculations). Values are means ± SD; n = 6 or 7. A and B: putative housekeeping genes. C and D: genes identified in this experiment to have shifted into polysomes after activation of PERK. E and F: genes reported in the literature whose mRNA shifted into polysomes after activation of PERK. Statistical analysis of the data is presented in Supplemental Table S8.