Supplementary Figure 1: Pilot CrY2H-seq experiments to confirm strain and plasmid functionality.
From: CrY2H-seq: a massively multiplexed assay for deep-coverage interactome mapping
(a) RT-PCR on HIS3 positive diploid cell lysate containing known interaction partners AT3G62420 (bZIP53) and AT5G28770 (bZIP63). Lanes 2-4, RT reaction with Cre primers, Lanes 8-10, RT reaction with Gal4 DBD primers (expression control). Control lane, genomic DNA from HIS3 positive diploid lysate. M, 100 bp DNA ladder. (b) Colony PCR on HIS3 positive diploids containing interacting pair bZIP53 and bZIP63 using primers to amplify Cre-recombined ORFs. Lanes 1,3,5 show lox plasmids in Y8800/CRY8930; lanes 2,4,6 show lox plasmids in Y8800/ Y8930; lane 7 shows two non-recombined lox plasmids (negative PCR control); lane 8 shows Cre-recombined ORF products (positive PCR control). (c) Colony PCR to amplify Cre-recombined ORFs from cells containing non-interacting pairs. Lanes 1,3,5 and 9,11,13 show pADlox-ZTL and pDBlox-bZIP63 in Y8800/CRY8930; lanes 2,4,6 and 10, 12,14 show pADlox-bZIP53 and pDBlox-ZTL in Y8800/CRY8930; lanes 7 and 15 show two non-recombined lox plasmids (negative PCR control); lanes 8 and 16 show Cre-recombined plasmid (positive PCR control).
