High-throughput measurement of DNAJB6b suppression of mHttex1 aggregation
(A) Fluorescence intensities and ratiometric images of Q71-VC with and without co-expression of DNAJB6b. Scale bars are 10 μm.
(B) FRET versus mVenus channel of Q71-VC, Q25-VC, co-transfection with DNAJB6b, and VC-DNAJB6b in HEK293T cells as obtained by FACS showing clear differences in populations. Biological replicates are shown in Figure S5.
(C) Violin plots of the individual FRET/donor ratios obtained by FACS in (B).
(D) Left: diagram illustrates the percentage of high-FRET cells at different Q71-VC expression levels when cells are co-transfected with different amounts of DNAJB6b plasmid. Right: density maps of FACS data show the distribution of cells in FRET-ratio versus protein-expression levels, demonstrating two separate groups of Q71-VC when Q71-VC is co-transfected without and with DNAJB6b plasmid (DNAJB6 plasmid amount corresponds to white line in the left graph). Cells were fixed with 4% paraformaldehyde (PFA). Cells were imaged 24 h after transfection.