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. 2019 Oct 16;218(12):4030–4041. doi: 10.1083/jcb.201811136

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© 2019 Wang et al.

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Figure 4. — Depletion of Rab7 inhibits cilia ectocytosis during cilia disassembly. (A) Knockdown of Rab7 inhibits cilia ectocytosis. RPE-1 cells stably expressing GFP-SMO were transfected with the indicated siRNA or treated with 200 nM LatA (positive control) during serum restimulation for live-cell imaging. Arrowheads, excised cilia tips. Time in hour:minute. Scale bar, 5 µm. (B) Scoring of the percentage of cells that underwent cilia ectocytosis under quiescent (−serum) or growth-stimulated (+serum) conditions. Data are means ± SD of three independent experiments. Student’s t test was performed. **P < 0.01. (C) The percentages of transfected cells that underwent cilia ectocytosis under growth-stimulated (+serum) conditions were quantified. Cilia ectocytosis defects induced by Rab7 knockdown in RPE-1 cells were rescued by mCherry-Rab7-WT or mCherry-Rab7-Q67L, but not by the T22N mutant of Rab7. Data are means ± SD of three independent experiments. Student’s t test was performed. **P < 0.01, ***P < 0.001. (D) Immunoblots of RPE-1 cell lysates in C with the indicated antibodies. GAPDH was used as a loading control.