Fig. 2: rNLS8 mice display dysregulation of genes in multiple cellular stress pathways and activation of the ISR in the cortex even prior to disease onset.

A Real-time qPCR verified upregulated RT² array-identified genes (Atf4, Chop, Bid, Trp53 and Gadd45g) in the cortex of rNLS8 mice at 2 and 4 weeks off Dox (WOD), and revealed similar upregulation at 1 WOD. B Additional ISR-related genes (Ppp1r15a and spliced Xbp1) were also upregulated at 1 WOD. C The RT² array-identified upregulated genes Ccl12, Il6, Tnf, Tlr4 and Homx1 were verified as increased at 2 and 4 WOD but were unaltered at 1 WOD. D Differential alterations were also identified in the RT² array-identified downregulated genes (Lhda, Slc9a3 and Crp) at 1 and 4 WOD. Values were normalized to Actb as the housekeeping gene. E Immunoblotting for the proteins of eIF2α and p-eIF2α (S51) in the cortex of rNLS8 mice at 4 WOD. Approximate molecular weights (kDa) are indicated. Quantification of immunoblots showed increased eIF2α (F) and p-eIF2α (G) in the cortex of rNLS8 mice relative to control mice, respectively. H Immunoblotting for ATF4 protein in the cortex of rNLS8 mice at 4 WOD. I Quantification of immunoblots showed increased ATF4 protein in the cortex of rNLS8 mice relative to control mice. n = 5. Mean ± SEM. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.