Figure 3.

Effect of trehalose on the remodeling of mHttt into forming IB is unaffected by knocking down of HSP70 or HSC70 expression. For HSP70 KD, cells were transfected either with control or HSP70 DsiRNA according to methods previously described⁹. For HSC70 KD, cells were transfected with the hsc70 antisense vector DNA as described⁹. 12 h after transfection and designated as t = 0 in the timeline of the experiment, cells were plated in a 96 well plate in medium containing 5 µM PA to induce the expression of mHtt. Trehalose was added at 24 h after plating and cells were harvested, fixed and processed for imaging at 48 h. (A) Images of control and trehalose (120 mM) treated cells without and with HSP70 or HSPC70 KD. (B) Bar graph on HSP70 or HSPC70 expression of cells under the specified conditions. (C) Quantitation of the total IB count/cell. (D) % distribution of mHtt in the IB versus diffuse format. (E) Timeline of experiment. Result shown is the average of results from three separate experiments with 15–20 images per condition. Statistical analysis was done using GraphPad Instat. Probability of difference P > 0.05 is defined as not significant, between 0.01 and 0.05 is significant (*), < 0.01 is very significant (**), and < 0.001 is extremely significant (***).