Fig. 6

The J-domain of DNAJB6b is critical for preventing tau aggregation. A Cellular lysate of SH-SY5Y cells co-transfected with mutated tau P301L together with an empty vector, V5-DNAJB6b, or V5-DNAJB6b H31Q plasmids for 48 h were filtered through cellulose acetate membranes. The dot blot assay was then performed with a tau antibody to detect trapped tau. B Quantification of tau aggregates is shown in A. The values were given as mean ± S.D. (n = 3, **P < 0.01, unpaired two-tailed Student’s t-test). C The expression levels of tau and DNAJB6b were determined by immunoblotting using tau and DNAJB6 antibodies, respectively. β-actin was used as a loading control for the normalization in B. D SH-SY5Y cells co-expressing tau P301L and an empty vector, V5-DNAJB6b or V5-DNAJB6b H31Q for 48 h were immunoprecipitated using an anti-V5 antibody to pull down DNAJB6b. Precipitated and co-precipitated proteins were analyzed by immunoblotting using indicated antibodies. E–F Quantification of the ratio of co-immunoprecipitated tau (E) and HSPA8 (F) to immunoprecipitated DNAJB6b or DNAJB6b H31Q in (D). The values were given as mean ± S.D. (n = 3, **P < 0.01, unpaired two-tailed Student’s t-test). The individual data values of the replicates in B, E, and F are listed in Additional file 2