Fig. 4.

UPR induction in cells expressing missense α-Gal A mutations. (A-C) Real-time RT-qPCR showing expression levels of HSPA5, XBP1s and DDIT3, normalized to HPRT1. Data are expressed as mean ± s.d. (n = 3 independent experiments) relative to wild type cells. (D) Western blot analysis of PERK in HEK293 GLA^−/− cells transfected with different α-Gal A isoforms. T indicates incubation with tunicamycin (2 μg/ml for 14 h), as positive control of PERK phosphorylation (molecular weight shift). (E) ATF6 activation assessed through the use of a luciferase-based, ATF6 reporter construct. Data are expressed as mean ± s.d. (n = 6 independent experiments). (A-E) All mutant isoforms were compared to wild type, one-way ANOVA followed by Dunnett's multiple comparison test. **p < 0.01, ***p < 0.001.