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. 2014 Oct 31;9(10):e111384. doi: 10.1371/journal.pone.0111384

Figure 3. ASPP2 scaffolds PP1 to dephosphorylate YAP.

Figure 3

(A) The importance of the ASPP2/PP1 interaction in dephosphorylating YAP was tested by transfecting the indicated constructs in HEK293 cells. Exogenous YAP was subsequently immunoprecipitated using an anti-myc mouse monoclonal antibody (9E10) and SDS-Page/immunoblotting was performed using the indicated antibodies. The bar graph represents the ratio between pYAP S127 and total YAP protein levels (n = 3; *: p<0.05; n.s.: non-significant). Error bars indicate standard deviation. (B) Lysates obtained from HEK293 cells transfected with the indicated constructs were analysed by SDS-PAGE/immunoblotting on a Phos-Tag gel. (C) The ability of ASPP2 (Y869A/Y874A)-V5 to dephosphorylate YAP was tested in HEK293 cells to investigate the importance of the YAP/ASPP2 interaction in this process. Following transfection with the indicated constructs in HEK293 cells, Exogenous YAP was immunoprecipitated using an anti-myc mouse monoclonal antibody (9E10) and SDS-Page/immunoblotting was performed using the indicated antibodies. (D) The ability of ASPP2, iASPP and ASPP1-V5 to interact with and dephosphorylate YAP was tested in HEK293 cells. Following transfection with the indicated constructs in HEK293 cells, Exogenous YAP was immunoprecipitated using an anti-myc mouse monoclonal antibody (9E10) and SDS-Page/immunoblotting was performed using the indicated antibodies.