Figure 2

Regulated expression of 72Q Htt-exon1 and aggregation formation. A: Addition of 500 nM RSL1 inducer to the culture medium leads to a time dependent increase in 72Q Htt-exon1 expression first detectable after 6 h of treatment (western blot of biological triplicates). B: Time-dependent disappearance of 72Q Htt-exon1 in HN10 cells upon removal of the RSL1 inducer from the culture medium. C: Dose dependent Htt expression as a function of RSL1 concentration in the culture medium of 25Q Htt-exon1, 72Q Htt-exon1, 25Q Htt-857 and 72Q Htt-857 cell lines (western blot quantification). D: AGERA blot of 72Q Htt-exon1 cell lysates reveals significant accumulation of Htt aggregates after induction of Htt expression but not in non-induced cells. E: Subcellular fractionation of cytoplasma and nuclei demonstrate the presence of abundant amounts of soluble Htt in the cytoplasma of the HN10 cells. In contrast, a minor pool of soluble mutant Htt and all aggregated 72Q Htt-exon1 were detected in the nuclei.