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High-pressure freezing reveals that ILVs may be connected by intramembranous material. Control or Hrs-depleted HeLa cells were incubated with 10 nm BSA-gold for 2 h at 37°C. Cells were stimulated with EGF and 5 nm anti-EGFR-gold for 25 min prior to fixation and then analysed by conventional EM. Endosomal clathrin coats are shown by arrows. Intramembranous fibrils are shown by arrowheads. Scale bar: 100 nm
