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. 2012 Sep 20;7(9):e45819. doi: 10.1371/journal.pone.0045819

Figure 1. GAGE proteins interact with GCL.

Figure 1

A. GAGE12I was used as bait in a yeast two-hybrid screen of a testis cDNA library. Plasmids encoding putative preys identified in the primary screen were rescued and retransformed into EGY42 with the bait GAGE12I vector (B) or control vector (C). Clone D2 exhibited growth on Ura- medium and blue coloration on X-Gal medium, indicating an interaction between bait and prey. The encoded prey polypeptide of D2 comprised human GCL residues 84–320, which includes the proposed BTB/POZ and BACK domains. B. Interaction between GCL and GAGE12I was verified by Lumier pull-downs from HEK293 cells co-transfected with Protein A-fused GAGE and Luciferase-fused GCL. Normalized luciferase signals (bound/input) are presented as Z scores, representing the number of standard deviations from the mean of a large set of independently derived, non-interacting Lumier protein pairs (see Materials and Methods for details). C. Pull-down assay with recombinant His-tagged GAGE12I and GST-GCL.