Figure 6. Enhanced IFT20 intensity induced by serum starvation at centrioles depends on Rab8 activation.

• A
  Efficiency of Rab8a depletion confirmed by Western blot in RPE1 cells.
• B
  RPE1 cells were transfected by siRNAs as indicated, followed by incubation in serum‐starvation media for 24 h, and immunostained for ARL13B (red) and γ‐tubulin (green). Scale bar, 10 μm.
• C
  Quantification of the percentage of ciliated cells shown in (B). Data represent mean ± SD (n = 3 experiments), and 250 GFP‐positive cells were scored per condition per experiment; *P < 0.05, Student's t‐test.
• D
  Cells were transfected by siRNA as indicated followed by incubation in serum‐starvation media for 24 h, and immunostained for EHD1 (red) and γ‐tubulin (green). Scale bar, 10 μm.
• E
  Quantification of the percentage of cells with EHD1 in cilium or in the distal end of basal body as shown in (D). Data represent mean ± SD (n = 3 experiments), and 150 cells were scored per condition per experiment; *P < 0.05, Student's t‐test.
• F
  Cells were transfected by siRNAs as indicated, followed by incubation in serum‐starvation media for 24 h, and immunostained for IFT20 (red) and γ‐tubulin (green). Scale bar, 10 μm.
• G
  Quantification of the percentage of IFT20 fluorescent intensity at the centriole shown in (F). Data represent mean ± SD (n = 3 experiments), and 150 were scored per condition per experiment; *P < 0.05, Student's t‐test.
• H–J
  Cells were transfected by siRNAs as indicated, followed by transfection with GFP‐Rab8 WT, GFP‐Rab8 Q67L, or GFP‐Rab8 T22N, and further incubated in serum‐starvation media for 12 h. Cell lysate was subjected to immunoprecipitation with anti‐GFP antibody, followed by Western blot with antibody against GFP.

Source data are available online for this figure.