Fig. 1

CapZ is transiently recruited to the early endosomes. A Live RAB5-GFP/CapZβ-mCherry-expressing HeLa cells treated with or without V1 (1 mM) were imaged using an N-SIM super-resolution microscope system (Movie S1 and S2), and selected frames of the time-lapse movies are presented here. The arrows in the upper panel indicate the transient association of CapZ puncta with RAB5-positive endosomes in control cells, and the dashed circle in the lower panel indicates the V1-induced tight association between CapZ and the enlarged RAB5-positive endosomes. Total 5 ~ 8 cells in either groups were imaged in 3 independent experiments. B–E Myc-CapZβ with Flag-Rab5 (B), HA-Rabex-5 (C), Flag-Rabaptin-5 (D), or Flag-Vps34 (E) were transiently transfected into HEK 293 T cells. The cell lysates were then incubated with anti-Myc-magnetic beads, and the pulldowns were subjected to immunoblot analysis against H.A.- or Flag-tag. The co-IP experiments were repeated three times independently. The difference between two groups was calculated using unpaired Student’s t-test. Differences were considered statistically significant when P < 0.05, *** P < 0.001