Figure 2 of Chen, Mol Vis 2008; 14:2076-2086.


Figure 2. Prox1 interacted with the IDCL and the carboxyl-terminal domain of PCNA. A: Diagram of the structure of full length PCNA and the structure of the truncations used in this study. N+ (amino acids 1-135 of PCNA); C+ (amino acids 115-261 of PCNA); C- (amino acids 135-261 of PCNA); C-term, the original clone from the yeast two hybrid screen (amino acids 167-261 of PCNA). B: Yeast strain AH109 transformed with a vector expressing a fusion between the DNA binding domain of GAL4 and the HDPD of Prox1 was mated with yeast strain Y187 transformed with a vectors expressing either the activation domain of GAL4 fused to the C-term fragment of PCNA (2), the C- fragment of PCNA lacking the IDCL (3), the C+ fragment of PCNA which includes the IDCL (4), the N+ fragment of PCNA (5) or full length PCNA (6). The interaction between P53 and SV40 large T-antigen was used as a positive control (1). Colonies from the original DO plates were re-streaked on either DO plates (to demonstrate presence of all vectors in the yeast) or quadruple drop out (QDO) plates to test for interaction between the protein products. C: QDO liquid cultures were grown of the yeast able to grow under QDO conditions and the relative amount of expression from the β-galactosidase reporter gene was determined by ONPG assays.