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. 2017 Oct 17;292(50):20628–20643. doi: 10.1074/jbc.M117.800946

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Figure 6. — Numb/Numblike knockdown impairs the interaction between Dll4 and AP1γ1. A, co-immunoprecipitation of Numb and HA-Dll4 from AD293-HA-Dll4 cell lysates. B, Dll4 contains (DE)XXXL(LI)-related motif and YXXØ motif in the cytoplasmic tail. C, AD293-HA-Dll4 cells transfected with or without FLAG-tagged AP1μ1 were treated with or without cross-linking. The lysates were then harvested and subjected to immunoprecipitation (IP) and immunoblotting as indicated. D, AD293 cells transfected with Myc-Numb72 were treated with or without cross-linking and then lysed for co-immunoprecipitation as indicated. E, control and Numb/Numblike knockdown cells (AD293-HA-Dll4) treated with cross-linking were lysed and subjected to immunoprecipitation and immunoblotting as indicated. F, AD293-HA-Dll4 cells transfected with indicated siRNAs were stained for surface HA-Dll4 and AP1γ1. Surface HA immunofluorescence intensity was quantified and shown in the right panel. Each data point is expressed as a percentage relative to that in control cells. (n = 20 cells for each condition, means ± S.D.) ***, p < 0.001.