Supplementary Figure 7: Uncropped gel images.
From: A molecular code for endosomal recycling of phosphorylated cargos by the SNX27–retromer complex
(a) Related to Fig. 3d . GFP-DGKζ wild type and mutant constructs expressed in Jurkat T-cells were analysed for binding to SNX27 by GFP-trap immunoprecipitation. (b) Related to Fig. 5c . GST-pulldown experiments from HEK293 cells transfected with myc-SNX27 and GST-GluN1 or GST-GluN2B C-terminal tails, either WT, PDZ mutant, phosphodefective (AA) or phosphomimetic (DD) mutants. (c) Related to Fig. 7b . Steady state levels of putative SNX27 cargos were reduced following SNX27 knockout by CRISPR-Cas9 deletion. Whole cell lysates from control HeLa cells (stably expressing Cas9) and SNX27 knockout cells were probed for several PDZbm-containing cargos by western blotting. Reduction of PDGFRβ, ATP7A and SNX14 suggests a defect in endosomal transport leading to lysosomal degradation. EAAT1 total levels were apparently unaltered. Na+/K+-ATPase is a control plasma membrane protein.
