Figure 5.

ERAD and UPR activation in GBA1b^m/m flies. (A) Protein lysates prepared from HEK293T cells, transfected with plasmids expressing normal or mutant GBA1b-encoded proteins (mycHispcDNAGBA1b-GBA1b, mycHispcDNAGBA1b^m-GBA1b^m), were treated or untreated for 18 h with MG132. The corresponding blot was interacted with anti-myc and anti-Erk antibodies. (B) To quantify the results, WT and mutant myc-GBA1b intensity was divided by that of Erk in the same lane, and the number obtained for GBA1b without treatment was considered 1. The results represent the mean ± standard error of three independent experiments. (C,D) mRNA levels of UPR markers: activating transcription factor 4 (ATF4), Heat shock-70-3 (HSC-70-3) and spliced x-box binding protein (sXBP1), in bodies (C) and heads (D) of control (w1118), and GBA1b^m/m flies at 2 and 18 days post-eclosion, as analyzed by qRT-PCR. mRNA level of each marker was normalized to that of RP49. mRNA levels of the different tested genes in control flies were considered 1. * p < 0.05, ** p < 0.01, *** p < 0.005.