Figure EV3. Immuno‐electron microscopy of damaged lysosomes and TAX1BP1 localization upon lysosomal damage (related to Fig 4).

• A–C
  Immuno‐electron microscopy of LLOMe‐treated HeLa cells. Cells in (A) and (B) overexpress dominant‐negative C160S mutant of GFP‐YOD1 and mCherry‐Gal3, and were immunostained for Gal3 and GFP. (A) GFP‐YOD1 (10 nm gold particles) colocalizes with mCherry‐Gal3 (15 nm gold particles) in the lumen of lysosomes (L). (B) mCherry‐Gal3 (10 nm gold particles) positive lysosomes (at the left) are morphologically indistinguishable from galectin‐negative lysosomes (right). (C) UBE2QL1‐GFP (10 nm gold particles) in late endosome/lysosomal compartments (LE). N = nucleus, M = mitochondrion. Scale bars: 200 nm.
• D
  HeLa cells were treated with LLOMe for 1 h. After methanol fixation, TAX1BP1 and LAMP1 were stained with specific antibodies and images were obtained by confocal microscopy. Note the recruitment of TAX1BP1 to lysosomes upon LLOMe treatment. Arrows indicate colocalizing vesicles. Scale bar: 20 μm and 2 μm for inlays.