(A and B) WT and myrlysin-KO HeLa cells were transiently transfected with plasmids encoding Myc-SKIP (A) or Lamp-1-KBS-GFP (B) and fixed 48 hr thereafter (KBS, KLC-binding sequence). Cells were immunostained for Myc, GFP, LAMTOR4, and/or CD63 and examined by confocal microscopy. Scale bar, 10 μm.
(C) WT and myrlysin-KO cells were transiently transfected with a plasmid encoding Lamp-1-GFP and imaged live 48 hr later. The images show the first frames from Movie S1 (left). Anterograde (red) and retrograde (green) movements were tracked using ImageJ over a 19-s period. The number and distance of lysosome movements were quantified from the movies for five cells in each group. Values are mean ± SD. The p values were calculated using Student’s t test (right). Scale bar, 5 μm.