Skip to main content
. 2005 Dec 2;33(21):6805–6815. doi: 10.1093/nar/gki985

Figure 2.

Figure 2

Analysis of the interaction between Glis2 and the co-repressor CtBP1 by mammalian two-hybrid and immuno-pulldown analysis. (A) Mammalian two-hybrid analysis. CHO cells were co-transfected with (UAS)5-LUC, phRL-SV40, pM, pVP16, pM-Glis2, and increasing amounts (µg) of pVP16-CtBP1 as indicated. Cells were assayed for reporter activity 48 h after transfection. The relative LUC activity was calculated and plotted. (B) Immuno-pulldown analysis. CV-1 cells were co-transfected with p3XFlag-CMV-Glis2, pCMV-myc-CtBP1, p3XFlag-CMV, and pCMV-myc as indicated in the graph. Cell lysates were prepared and Flag-Glis2 protein complexes isolated using anti-Flag M2 agarose affinity resin as described in Materials and Methods. Proteins in the cellular lysates and immunoprecipitated (IP) proteins were subsequently examined by western blot (WB) analysis using anti-Flag M2 or anti-myc antibody. (C) Glis2 interacts with CtBP1 in different cell types. CHO, CV-1, 293HT and COS-1 cells were co-transfected with (UAS)5-LUC, phRL-SV40, pM, pVP16, pM-Glis2, pVP16 and pVP16-CtBP1 as indicated. Cells were assayed for reporter activity 48 h after transfection and the relative LUC activities calculated and plotted.