Extended Data Fig. 7. Intramolecular transmembrane segment interactions are crucial for seipin function.
(a) Western blot analysis of whole-cell lysates from strains expressing C-terminal 13xmyc tagged seipin variants from endogenous or PGK1 promoter. (b) Western blot analysis of fractions from size-exclusion chromatography of Triton X-100 solubilized membrane extracts carrying Patch mutants combined with R178A and C-terminal 13xmyc. (c) Growth of yeast strain sei1∆ carrying plasmids with C-terminally GFP-tagged SEI1 from yeast (WT), or indicated mutants. (d) Western blot analysis of whole-cell lysates from strains expressing indicated seipin mutants under control of the PGK1 promoter and C-terminal 13xmyc tag. Representative immunoblots of two biologically independent experiment repeats is shown. (e) Immuno-precipitation of indicated seipin mutants via anti-myc resin. Equal amounts of load (detergent solubilized membranes in Tx100) and eluate fractions were loaded. n=2 biologically independent repeats.