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. 2021 Nov 23;221(1):e202105092. doi: 10.1083/jcb.202105092

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© 2021 Shen et al.

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Figure 6. — PRPF8 colocalizes with CP110 and disappears from the distal end of the mother centriole in the initiation of ciliogenesis. (A) RPE-1 cells were transfected with mCherry-Centrin2 for 24 h. The cells were stained with the indicated antibodies. PRPF8 was visualized with centriolar proximal (C-Nap1), distal (Centrin2), subdistal appendage (ODF2), and distal appendage (CEP164) markers. Scale bar, 0.5 µm. (B) PRPF8 was colocalized with CP110. RPE-1 cells were transfected with GFP-CP110 for 24 h. The cells were stained with the indicated antibodies. Scale bar, 0.5 µm. (C) The relation between the relative position and fluorescence intensity of PRPF8, CP110, and CEP164 in B. (D and E) RPE-1 cells were serum starved for the indicated time points and stained with the indicated antibodies. Scale bars, 1 µm. Immunostaining of PRPF8 and CP110 are shown in D and E, respectively. (F) Line graph showing the percentages of cells with PRPF8 and CP110 on the mother centrioles in D and E. (G) RPE-1 cells were transfected with control or HOIP siRNAs and serum starved for the indicated time points. The cells were collected and stained with the indicated antibodies. Scale bar, 1 µm. (H) Quantification of cells with centriolar PRPF8 in G. Data are presented as mean ± SD of three independent experiments. Ctrl, control.