Figure 1. Members of a DNAJB Subfamily Act as Strong Suppressors of Polyglutamine Aggregation (A) Cells were cotransfected with HDQ119-EYFP and V5-tagged members of the HSPH, HSPA, or DNAJ families. Expression of chaperones was induced with tetracycline. Samples were taken 24 hr after transfection. High-molecular-weight (HMW) aggregates trapped in the stacking gel and soluble HDQ119-EYFP were assessed with anti-GFP antibodies. Expression of chaperones was detected with anti-V5 antibodies. (B) Filter trap assay of cell extracts without and with overexpression of molecular chaperones. Serial 5-fold dilutions were loaded on cellulose acetate membranes and probed with anti-GFP antibody. Numbers represent the percentage of aggregation in cells with exogenous chaperones (+Tet) compared to cells without the exogenous chaperones (−Tet). (C) Representative confocal pictures of cells cotransfected with HDQ119-EYFP (green) and V5-tagged chaperone members (red). DAPI staining is shown in blue. Bar represents 30 μm. (D) Combined phylogram of aligned DNAJB primary amino acid sequences (left) with aggregation suppressive activity in the filter trap assay (right; efficacy indicated in percentages as above). Bootstrap values of 1000 trials on each node are given.
Image published in: Hageman J et al. (2010)
Copyright © 2010. Image reproduced with permission of the Publisher, Elsevier B. V.
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