Figure 6.

CHOP function is not altered in S63del//Perk+/− mice. (a) WB analysis on P28 sciatic nerves for ATF4; protein levels were quantified by densitometric analysis. One representative blot of three independent experiments is shown. Error bars, SEM; n = 3. Loading control, beta tubulin. (b) Quantitative RT-PCR for Chop mRNA from WT, Perk+/−, S63del, and S63del//Perk+/− sciatic nerves at P28. Error bars, SEM; ***p < .001, by Student’s t test; n = 3 animals for each genotype. (c) Confocal immunofluorescence for CHOP (red) on transverse sections of P28 sciatic nerves. MBP (green) identifies myelinating cells; nuclei are visualized with DAPI (blue). Arrowheads indicate CHOP-positive nuclei. A typical image of one out of three independent experiments on separate animals is shown. As controls, S63del-H (210% overexpression), and Chop−/− showed strongly positive and negative signals, respectively (not shown). Bar: 50 microns. (d) Quantitative RT-PCR for Gadd34 mRNA from WT, Perk+/−, S63del and S63del//Perk+/− sciatic nerves at P28. Error bars, SEM; *p < .05; Student’s t test; ns, nonsignificant; n = 3 independent replicates. (e) WB analysis on P28 sciatic nerves was performed for Gadd34; protein levels were quantified by densitometric analysis. β-Tub was used as loading control. One representative blot of seven independent experiments is shown. Error bars, SEM; n = 7. Numbers represent relative molecular weights. DAPI = 4′,6-diamidino-2-phenylindole; MBP = myelin basic.