Interactions of Sec61p and Ssh1p bait proteins using PLV cleavage as readout. Prey proteins were expressed in heterozygous diploids expressing the bait-fusion proteins as above. Cells were grown to early exponential phase in minimal medium, and lysates of 1 OD600 were separated by SDS-PAGE, proteins transferred to nitrocellulose, and quantitative immunoblotting performed with polyclonal rabbit antibodies against PLV, to detect intact and cleaved bait proteins, and Arf1p (as loading control) and 125I-Protein A. The ratios of cleaved PLV to Arf1p for each strain are shown in the graph in blue for Sec61-CPLV and yellow for Ssh1-CPLV.
Harty and Römisch BMC Cell Biology 2013 14:14 doi:10.1186/1471-2121-14-14