ERdj8 knockdown allows enwrapping of only smaller target. (A) HeLa cells were treated with siRNA (siERdj8no1 or control), incubated for 96 h, and then treated with 5 µg/ml puromycin for 6 h. After washout, cells were cultured for 17 h, stained with anti-p62, and imaged on an SP-8. Scale bar, 10 µm. (B) Percentage of numbers of p62-positive puncta longer than 2 µm in a cell after washout versus before washout. *, P < 0.05 by ANOVA, Tukey Kramer test. NS means P > 0.05. Median: lines; upper and lower quartiles: boxes; 1.5-interquartile range: whiskers. (C) HeLa subjected to the indicated siRNA or ATG16L-knockout stably expressing mRFP-eGFP-Galectin8 was incubated with 3-µm beads for 24 h and imaged on a DeltaVision system. Scale bar, 10 µm. Insets, enlargements of framed regions. (D) GFP and RFP signal intensities associated with 3-µm beads were measured; the GFP/RFP ratio of 61 beads is shown. ***, P < 0.001. NS means P > 0.05. Median: line; upper and lower quartiles: boxes; 1.5-interquartile range: whiskers.