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. 2017 Jul 20;23(5):1278–1286. doi: 10.1038/mp.2017.138

Figure 2.

Figure 2

FBXW7 binds DISC1 via a PD motif to regulate turnover of DISC1. (a) HEK293 cells were transfected with FLAG-tagged FBXW7 or FLAG-tagged FBXW7 without the F-Box substrate-binding domain (Δ-F). Lysates were prepared and blotted for DISC1, c-MYC, FLAG and GAPDH. Relative levels of DISC1 expression were calculated (right hand panel). *=P<0.05 as measured by Student’s t-test, n=3 (b) HEK293 cells were transfected with FLAG-FBXW7 and GFP-DISC1 (upper panels) or FLAG-FBXW7 and V5-DISC1 (lower panels). FLAG was immunoprecipitated from cell lysates and blotted for FLAG, GFP and V5. (c) HEK293 cells were transfected with FLAG-DISC1. FLAG was immunoprecipitated from cell lysates and blotted for FLAG and endogenous FBXW7. (d) and (e) Peptide arrays of sequences containing the DISC1 PD were overlayed with a purified protein complex containing 6His-FBXW7 and Skp1. Arrays were then probed for His tag. (f) Sequences of known FBXW7 substrates contain a PD that is also present in DISC1.