Supplementary Figure 5: Gel analysis of cytoplasmic dynein constructs.

(a) SDS-PAGE analysis (4–15%) of recombinant full-length human and native porcine dyneins. Both the human and porcine dynein heavy chain were co-purified with endogenous intermediate chains (ICs), light intermediate chains (LICs) and light chains (LCs), but without the dynactin complex. The protein bands were visualised by Stain-Free technology (Bio-Rad). (b) SDS-PAGE analysis (4–15%) of the GST-D425 dynein. The image shows the absence of subcomponents in the GST-D425 dynein. The protein band was visualised by Stain-Free technology (Bio-Rad). (c) SDS-PAGE analysis of two coupled motor domains of dynein on DNA scaffolds with the spacing of 3 nm, 17 nm and 32 nm. SDS-PAGE were performed on a 4% polyacrylamide gel. After electrophoresis, the gel was stained with Coomassie Brilliant Blue G250-based reagent (24590, Thermo Scientific). (d) SDS-PAGE analysis of two coupled full-length dynein molecules. SDS-PAGE was performed on a 5% polyacrylamide gel. (e) Gel shift assay of Alexa Fluor 647-labelled DNA origami nanotubes carrying 4 and 8 dynein molecules. Electrophoresis was performed on a 0.5% agarose gel. The DNA bands were visualised by epifluorescence imaging with red LED excitation (ChemiDoc MP, Bio-Rad).