Extended Data Fig. 2: Biochemical properties of IST2716 and lipid transport experiments.
From: Structural basis for lipid transport at membrane contact sites by the IST2–OSH6 complex
a, Size exclusion chromatogram of a sample of IST2716 and nhTMEM16 used for liposome reconstitution. b, Western Blots against n-terminal myc tag of nhTMEM16 and IST2716 reconstituted into liposomes for scrambling assays (n = 3, all shown). c, Scheme of the lipid scrambling assay. ‘red.’ refers to the addition of dithionite to the outside of liposomes. Fluorescent lipids are indicated as yellow, lipids with bleached fluorophore as grey spheres, a scramblase mediating the bidirectional diffusion of lipids between leaflets as cyan box. d-f, Scrambling data from three independent reconstitutions that are combined in Fig. 1f, g. The displayed nhTMEM16 and IST2716 preparations were reconstituted into the same batch of destabilized liposomes. Empty liposomes of the same batch are shown as negative control (neg.). Recordings show mean of three technical replicates, errors are s.d. g-h, Size exclusion chromatograms and SDS PAGE (inset, n = 1) of IST2716 in detergent (g) and after reconstitution in lipid nanodiscs (h) used as samples for the cryo-EM datasets shown in Extended Data Figs. 3 and 4, respectively. Dashed lines indicate pooled fractions.
