Extended Data Figure 7: Defining Get1/2 transmembrane Cys positions that interact with tail-anchored proteins.
From: The Get1/2 transmembrane complex is an endoplasmic-reticulum membrane protein insertase
a, Cells expressing the indicated single-cysteine alleles of Get1 and Get2 from their endogenous genetic loci and the indicated control cells were subjected to heat-shock reporter analysis as in Fig. 1a (n = 2). b, Cell extracts containing Get3–Sec22S192C were incubated with S protein followed by the addition of the indicated microsomes for 15 min at room temperature and crosslinking analysis as in Fig. 3c. Samples were denatured with SDS (an ionic detergent) and diluted into immunoprecipitation buffer with Triton X-100 (a non-ionic detergent) before pull-down with anti-Flag resin. Eluted material was subjected to SDS–PAGE analysis and visualized by immunoblotting and autoradiography. c, Get3–Sec22S192Cs was road-blocked with S protein and incubated with the indicated microsomes followed by crosslinking analysis as in Fig. 3c.
