Figure 4.
Figure 4. Rab8 is necessary for the movement of the midbody remnant. (A–D) Control cells (A and B) or cells stably expressing cherry-Rab8 (C and D) were transfected with siRNA control (siC) or targeted to Rab8 (siRab8). (A and C) Cells were stained for α- and γ-tubulin (α- and γ-tub). The green and red arrowheads point to the midbody remnant and the centrosome, respectively. The white arrowheads indicate the PCs. The asterisks in C mark the cells expressing cherry-Rab8. (B) The number of cells with a peripheral remnant or a PC was quantified in Rab8-knockdown cells and was expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 381 control cells and 354 Rab8-knockdown cells; two to three fields per experiment; Student’s t test). (D) The number of cells with a peripheral remnant or a PC was quantified in siRab8-transfected cells expressing cherry-Rab8 and was expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 294 control cells and 322 Rab8-knockdown cells; three fields per experiment; Student’s t test). (E and F) Cells were transfected with siC or siIFT88. (E) Cells were stained for α- and γ-tubulin. The green and white arrowheads point to the midbody remnant and the PC, respectively. (F) The number of cells with a peripheral remnant or a PC in IFT88-knockdown cells is expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 1,201 control cells and n = 1,329 IFT88-knockdown cells; three fields per experiment; Student’s t test). (G) Panoramic EM image of an apical zone with a midbody remnant in a Rab8-knockdown cell (top) and enlargements of serial sections of the remnant region from the same cell. The sections were numbered S1 onwards from the bottom to the top. Panels S2′ and S3′ show an enlargement of the boxed regions in S2 and S3, respectively. Note that it appears that the remnant is connected to the rest of the cell by a thin tether, indicated by an empty arrowhead. Bars: (A, C, and E) 5 µm; (G, panoramic view and S2–S12) 1 µm; (G, S2′ and S3′) 200 nm.

Rab8 is necessary for the movement of the midbody remnant. (A–D) Control cells (A and B) or cells stably expressing cherry-Rab8 (C and D) were transfected with siRNA control (siC) or targeted to Rab8 (siRab8). (A and C) Cells were stained for α- and γ-tubulin (α- and γ-tub). The green and red arrowheads point to the midbody remnant and the centrosome, respectively. The white arrowheads indicate the PCs. The asterisks in C mark the cells expressing cherry-Rab8. (B) The number of cells with a peripheral remnant or a PC was quantified in Rab8-knockdown cells and was expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 381 control cells and 354 Rab8-knockdown cells; two to three fields per experiment; Student’s t test). (D) The number of cells with a peripheral remnant or a PC was quantified in siRab8-transfected cells expressing cherry-Rab8 and was expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 294 control cells and 322 Rab8-knockdown cells; three fields per experiment; Student’s t test). (E and F) Cells were transfected with siC or siIFT88. (E) Cells were stained for α- and γ-tubulin. The green and white arrowheads point to the midbody remnant and the PC, respectively. (F) The number of cells with a peripheral remnant or a PC in IFT88-knockdown cells is expressed relative to that in siC-transfected cells. Data represent the mean + SEM from three independent experiments (n = 1,201 control cells and n = 1,329 IFT88-knockdown cells; three fields per experiment; Student’s t test). (G) Panoramic EM image of an apical zone with a midbody remnant in a Rab8-knockdown cell (top) and enlargements of serial sections of the remnant region from the same cell. The sections were numbered S1 onwards from the bottom to the top. Panels S2′ and S3′ show an enlargement of the boxed regions in S2 and S3, respectively. Note that it appears that the remnant is connected to the rest of the cell by a thin tether, indicated by an empty arrowhead. Bars: (A, C, and E) 5 µm; (G, panoramic view and S2–S12) 1 µm; (G, S2′ and S3′) 200 nm.

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