Extended Data Fig. 4: Additional characterization of substrate features that direct assembly of the multipass translocon.
From: Substrate-driven assembly of a translocon for multipass membrane proteins
a, The indicated ASGR1-derived intermediates were translated in RRL in the presence of wild-type HEK293 rough microsomes, and the membrane-associated fraction was isolated by sedimentation. Following anti-Flag immunoprecipitation of the digitonin-solubilized membranes, stalled RNCs were isolated by sedimentation and analyzed by SDS-PAGE and immunoblotting. b, Quantification of STT3A, TMCO1, CCDC47 and NOMO recruitment to stalled 2-TMD YIPF1 constructs as in Fig. 2f, for n = 5 biological replicates. Data are mean ± S.D. Analysis was performed using an RM one-way ANOVA with the Geisser-Greenhouse correction and Tukey’s multiple comparisons test as implemented in GraphPad Prism 9.4.0. *, p < 0.0332; **, p < 0.0021; ***, p < 0.0002; ****, p < 0.0001; p values are given in Supplementary Table 2.
