Fig. 9.

Effects of the c.695_699del recessive mutation in DNAJB6 on its ability to prevent aggregation of polyQ in cells. a Representative immunofluorescent images of HEK293 cells cotransfected with HttEx1-(Q)74-hrGFP and 500 ng empty vector as control (Con), 500 ng wild type M2-fused DNAJB6a (WT), 200 ng, 500 ng or 1500 ng mutant M2-fused DNAJB6a (200 ng mut, 500 ng mut, 1500 ng mut) plasmids respectively for 48 h. Scale bar = 50 µm. b Western blot showing Lysates from cells treated as in a. with antibodies against GFP, M2, and β-actin. c Quantitative analysis of a, expressing the proportion of cells containing inclusions (each scored by at least three independent observations and then averaged). d Filter-trap assay. Cell lysates in a. were loaded onto cellulose acetate membranes and probed with an anti-GFP antibody to detect aggregation of HttEx1-(Q) 74-hrGFP. e Quantification of data in d. Data from three experiments (mean ± SEM). P value = * < 0.05, ** < 0.01. N.S. = not significant