Expression of the mutant htt. (A) Quantitative analysis of htt transcripts in rHD/G-DPSCs and rG-DPSCs. qRT-PCR was used to measure the amount of htt transcripts. In addition to measuring the expression level of exon1, which is composed of both endogenous htt and the mutant htt, an additional set of primers measuring the expression level of Exon10/12 was used to show the level of normal htt alleles. The htt expression was significantly increased in all rHD-DPSC lines compared to rGFP. Column shares the same alphabet are significantly different (P < 0.05). (B) Western analysis demonstrated the accumulation of oligomeric htt (arrow) at high molecular weight (> 250 kD) that can be clearly seen in upper portion (Arrow) of a gradient polyacrylamide gel (upper panel). Antibody to γ-tubulin (bottom panel). (C) Subcellular distribution of mutant htt in rHD/G-DPSCs. DPSCs of rHD11, rHD17, rHD18 and rGFP were immunostained with mEM48. Transgenic mutant htt aggregates (arrows) and nuclear inclusions (arrowheads) were only observed in rHD11, rHD17 and rHD18 but not in DPSCs of rGFP. Scale bar = 5 μm.
Snyder et al. BMC Cell Biology 2011 12:39 doi:10.1186/1471-2121-12-39