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. 2018 Aug 22;46(21):11202–11213. doi: 10.1093/nar/gky754

Figure 1.

Figure 1.

DYRK1A co-purifies with p300 and CBP. (A) A clonal HEK293 cell line inducibly expressing Flag-DYRK1A was generated, and nuclear extracts prepared followed by affinity purification. Eluates were analysed by SDS-PAGE and mass spectrometry. Arrow marks the location of DYRK1A in the silver stained gel. (B) MudPIT analyses of the relative abundance of DYRK1A and highly enriched proteins are shown. The distributed Normalized Spectral Abundance Factor (dNSAF) reflects the relative abundance of the identified proteins in the samples (44). Many of the previously identified DYRK1A interacting proteins were reproducibly identified, including DCAF7 and ARIP4. These interactions were also observed with the kinase-dead DYRK1A-KD mutant. (C) Flag affinity pull-down of Flag-DYRK1A and 293T control whole cell extracts were probed with CBP and p300 antibodies, confirming interaction between DYRK1A and CBP/p300. (D) Flag affinity pull-down of Flag-DYRK1A-WT, Flag-DYRK1A-KD and 293T control whole cell extracts were probed with CBP and p300 antibodies, to compare the interaction efficiency of DYRK1A-WT and DYRK1A-KD with CBP/p300.