Extended Data Fig. 5: AUTAB induces membrane damage at cell surface.
From: Chemically engineered antibodies for autophagy-based receptor degradation
a, Over-expressing HA-PD-L1 and mCherry-LC3C HeLa cells were treated with 100 nM Atz or Atz-AUTAB for 1 h, then fixed and co-stained with antibodies against ATG9A and Atz. Images were taken by confocal microscopy. The white boxed regions in the images are enlarged below and the arrowheads point to the colocalization between ATG9A, mCherry-LC3C and Atz-AUTAB. The corresponding colocalization quantization is on the right (n = 15 cells per group, means ± s.d.). b, Plasma membrane permeabilization was evaluated with PI uptake assay in stably expressing HA-PD-L1 HeLa cells treated with 250 nM L25K PEI, 100 nM Atz, 25 nM and 100 nM Atz-AUTAB for 1 h. Percentage of cells with PI-positive was quantified (n = 3 biological replicates, means ± s.d.). DNA in b was counterstained with Hoechst 33258. Scale bars, 10 μm (a) or 100 μm (b). Statistical significance was calculated via unpaired two-tailed Student’s t-test (a, b).
