SNX17 depletion decreases Itgα5β1 recycling and leads to enhanced degradation of integrins caused by aberrant trafficking to lysosomes. (A) Itgα5β1 levels in control and SNX17-depleted HeLa cells treated with cycloheximide (Cyclohex.) over the indicated periods. Error bars indicate the standard deviation of three experiments. (B) Biochemical internalization assay of Itgα5β1 in control and SNX17-depleted cells. (C) Biotinylation-based recycling assay of Itgα5β1 in SNX17-depleted HeLa cells. The blot shows a representative experiment. (B and C) Error bars represent the SEM of four experiments. (D) Recycling assay with antibody against inactive (P5D2) Itgβ1. (E) Endosomal marker analysis of inactive Itgβ1 in vesicles remaining within the SNX17-depleted cells after a 30-min chase period. (F) Colocalization analysis of Itgβ1 and the lysosomal marker LAMP1 in control and SNX17-depleted cells treated with bafilomycin for 14 h. The Pearson’s correlation and the percentage of colocalization are based on eight images from two independent experiments. The error bars represent the standard deviation. Bars, 10 µm. IB, immunoblot; KD, knockdown.