Figure 2.
TBC1D20 regulates the localization of ciliary proteins. (A and B) Immunofluorescence images (A) and quantification of the percentage of cells with one CP110 dot (B) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against CP110 and γ-tubulin. n = 3 independent experiments. (C and D) Immunofluorescence images (C) and quantification of the percentage of cells with MKS3 at the centrosome (D) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against MKS3 and γ-tubulin. n = 3 independent experiments. (E and F) Immunofluorescence images (E) and quantification of the fluorescence intensity of IFT88 at the centrosome (F) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against IFT88 and CEP164. n = 3 independent experiments. (G and H) Immunofluorescence images (G) and quantification of the fluorescence intensity of IFT140 at the centrosome (H) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against IFT140 and CEP164. n = 3 independent experiments. (I and J) Immunofluorescence images (I) and quantification of the fluorescence intensity of PCNT (J) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with antibody against PCNT. Bottom panels depict three-dimensional surface plot of the intensities portrayed in top panels. n > 30 cells. (K–M) Immunofluorescence images (K), the distribution area of PCM1 (L), and the quantification of the fluorescence intensity of PCM1 (M) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with antibody against PCM1. Bottom panels depict three-dimensional surface plot of the intensities portrayed in top panels. n > 30 cells. (N and O) Immunofluorescence images of the fluorescence intensity of IFT88 and IFT140 at the centrosome for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured without serum medium, and stained with the antibodies against IFT88 and CEP164. All experiments were repeated at least three times. Data are presented as mean ± SEM. ***P < 0.001, ****P < 0.0001; ns, not significant. Scale bar, 5 µm. Refer to the image caption for details.

TBC1D20 regulates the localization of ciliary proteins. (A and B) Immunofluorescence images (A) and quantification of the percentage of cells with one CP110 dot (B) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against CP110 and γ-tubulin. n = 3 independent experiments. (C and D) Immunofluorescence images (C) and quantification of the percentage of cells with MKS3 at the centrosome (D) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against MKS3 and γ-tubulin. n = 3 independent experiments. (E and F) Immunofluorescence images (E) and quantification of the fluorescence intensity of IFT88 at the centrosome (F) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against IFT88 and CEP164. n = 3 independent experiments. (G and H) Immunofluorescence images (G) and quantification of the fluorescence intensity of IFT140 at the centrosome (H) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with the antibodies against IFT140 and CEP164. n = 3 independent experiments. (I and J) Immunofluorescence images (I) and quantification of the fluorescence intensity of PCNT (J) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with antibody against PCNT. Bottom panels depict three-dimensional surface plot of the intensities portrayed in top panels. n > 30 cells. (K–M) Immunofluorescence images (K), the distribution area of PCM1 (L), and the quantification of the fluorescence intensity of PCM1 (M) for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured in serum medium, and stained with antibody against PCM1. Bottom panels depict three-dimensional surface plot of the intensities portrayed in top panels. n > 30 cells. (N and O) Immunofluorescence images of the fluorescence intensity of IFT88 and IFT140 at the centrosome for RPE1 cells transfected with control or TBC1D20 siRNAs, cultured without serum medium, and stained with the antibodies against IFT88 and CEP164. All experiments were repeated at least three times. Data are presented as mean ± SEM. ***P < 0.001, ****P < 0.0001; ns, not significant. Scale bar, 5 µm.

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