Fig. 5

TMEM55B/JIP4 drive retrograde lysosomal trafficking independent of RILP. a HeLa cells treated with JIP4 or Control siRNA were immunostained with antibody against LAMP-1. b Quantification of lysosome distribution shown as the percentage of total fluorescence signal detected at 0–5 μm, 5–15 μm, or >15 μm from nuclear rim. Quantified results are presented as mean ± s.e.m. using two-tailed t-test ^∗ P < 0.05, ^∗∗ P < 0.005 were considered significant, n ≥ 30. c Immunoblot of HeLa cells depleted of JIP4 or Control RNAi. d HeLa cells treated with JIP4 or Control siRNA were infected with adenovirus expressing GFP-TMEM55B for 24 h. Cells were fixed, permeabilized, and immunostained with antibodies against LAMP-1. Arrows denote bulk of lysosomal accumulation. e Quantification of lysosome distribution, percentage of total fluorescence signal detected at 0–5 μm or >15 μm from nuclear rim. Quantified results are presented as mean ± s.e.m. using two-tailed t-test ^∗ P < 0.05, ^∗∗ P < 0.005 were considered significant, n ≥ 10. f HeLa cells depleted of TMEM55B, JIP4, or expressing control RNAis were transfected with GFP-RILP for 24 h. Cells were fixed, permeabilized, and immunostained with antibodies against LAMP-1. Arrows denote bulk of lysosomal accumulation. Scale bars, 20 μm