Fig 1: Rab4- and Rab25-specific machinery is required for cell migration in the 3D matrix. (A–D) A2780-DNA3 and A2780-Rab25 cells were depleted of CLINT1 (A,B) or SH3BP5L (C,D) by siRNA and seeded into CDM for 4 h, before migration was analysed by brightfield time-lapse imaging for >16 h. Cell speed was analysed by manual tracking (A,C). Representative images from at least three independent experiments are shown (B,D). Red lines indicate the migration of cells marked with asterisks. Scale bars: 100 µm. (E,F) BT20 cells were depleted of Rab4a and Rab4b (indicated as Rab4ab), Rab11a and Rab11b (indicated as Rab11ab), or Rab25 by siRNA and migration analysis performed as above to determine cell speed (E) and cell persistence (F). (G) BT20 cells were depleted of CLINT1 or SH3BP5L by siRNA and migration analysis performed as above. (H) Representative images of BT20 cells analysed in G. Yellow lines indicate the migration of cells marked with asterisks. Scale bars: 50 µm. For A–H, n=90 cells from three independent experiments. Box plots are presented as described as in Fig. 2. Statistical analysis was performed with Kruskal–Wallis and Dunn's multiple comparisons test. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.