Fig. 2

Visualization of DNAJB6-dependent tau aggregation in neuroblastoma cells. A Schematic representation of BiFC maturation upon tau aggregation. Tau P301L was fused to one of the halves of the Venus protein (VN- or VC-). Aggregation of tau protein promotes the complementation of two non-fluorescent Venus halves and leads to the emission of fluorescence. B Representative fluorescent images of BiFC and DNAJB6. shLuc or shDNAJB6 SH-SY5Y cells was co-transfected with the BiFC-tau P301L constructs. The fluorescence emitted by Venus complementation was recorded 48 h after transfection. Cells were immunostained with a DNAJB6 antibody. Nuclei were counterstained with DAPI. Enlarged Venus images of the representative cells corresponding to the selected regions are shown. Scale bar: 20 μm. C Immunoblotting with anti-tau antibodies indicated the expression level of BiFC-tau P301L in SH-SY5Y cells. The immunoblotting of anti-DNAJB6 antibodies determined the knockdown of DNAJB6. D Quantification of BiFC-fluorescence of B. The percentage of fluorescent (BiFC positive) SH-SY5Y cells to the total number of cells was quantified. The values were given as mean ± S.D. (n = 3, *P < 0.05, ***P < 0.001, unpaired two-tailed Student’s t-test). The individual data values of the replicates in D are listed in Additional file 2