Skip to main content
. 2016 Aug 17;6:31665. doi: 10.1038/srep31665

Figure 4. The γ-Pcdh-C3 VCD interacts with Axin1.

Figure 4

(a) Lysates of HEK293 cells transfected with either GFP (control), FL-A3GFP, FL-B1GFP or FL-C3GFP were immunoprecipitated with anti-GFP and blotted for endogenous (human) Axin1. Axin1 co-immunoprecipitated with γ-Pcdh-C3 but not –A3 or –B1. All proteins encoded by the transfected constructs were present in the cell lysates used (right; input) and re-probing of blots with anti-GFP antibody confirmed immunoprecipitation. (b) Lysates from HEK293 cells co-transfected with GFP or FLC3-GFP and either FLAG-tagged zebrafish (Danio rerio) Axin1 (FLAG-drAxin1), Myc-tagged Xenopus Axin1 (Myc-xAxin1) or HA-tagged mouse Axin1 (HA-mAxin1) were immunoprecipitated with anti-GFP and blotted for the tagged Axin1. FL-C3GFP co-immunoprecipitated all three Axin1 orthologues. (c) Co-immunoprecipitation of endogenous HEK293 Axin1 as in (a) was performed using GFP, FLC3-GFP, C3VCD-GFP, NLSC3VCD-GFP and PalmC3VCD-GFP. In all cases, the VCD is sufficient to pull down endogenous Axin1; co-immunoprecipitation was more robust for PalmC3VCD-GFP. Blots shown are representative of at least 3 experiments. MW, molecular weight; kDa, kilodaltons.