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. 2007 Nov 5;35(21):e142. doi: 10.1093/nar/gkm932

Figure 2.

Figure 2.

Yeast two-hybrid interaction between pVHL and HIF1α. (a) CG1945BC (+ELB/C) or CG1945 (–ELB/C) yeast cells transformed with the indicated combinations of bait (pBridge-) and prey (pACT2-) plasmids were diluted to an OD600 nm of 0.2 and spotted on SC-Leu-Trp-Met (—LWM) control plates or SC-Leu-Trp-Met-His (—LWMH) reporter plates containing 1 mM 3-aminotriazole (3-AT). (b)–(f) Expression levels of components of the yeast two-hybrid system. Yeast cells were directly taken from the SC-Leu-Trp-Met control plates shown in (a), dissolved in sterile water and diluted to identical OD600 nm values. Protein expression levels were determined by western blots (WB) using antibodies against pVHL (b), HIF1α (c), PHD1 (d), PHD2 (e) and PHD3 (f). Shown are representative sets of samples taken from CG1945BC cells expressing the indicated pBridge (b, d–f) and pACT2 (c) constructs. Expression levels were independent of the presence of ELB/ELC and of specific combinations of bait and prey plasmids, as tested in random samples obtained from several different yeast colonies on the SC-Leu-Trp-Met control plates (data not shown). In (c), an asterisk indicates the HIF1α band.