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. 2006 Sep;17(9):3781–3792. doi: 10.1091/mbc.E06-02-0133

Figure 6.

Figure 6.

Moderate IFT20 knockdown reduces ciliary polycystin-2 levels. Rat kidney NRK cells lines with reduced IFT20 or IFT88 were generated by expression of shRNA constructs. (A) Western blot analysis showing the reduction of IFT20 and IFT88 in the two cell lines. TIM23 was used as a loading control. (B) Immunofluorescence images showing that cilia (arrows) are still formed in these cells (acetylated tubulin, red). IFT20 (top row) and IFT88 (middle row) are reduced in the respective knockdown line but not greatly altered in the reciprocal line. Ciliary polycystin-2 (PKD2, green) is reduced in the IFT20 knockdown line (bottom middle) compared with the control cells (bottom left) and the IFT88 knockdown line (bottom right). The insets in the bottom row show ciliary polycystin-2 staining without acetylated tubulin staining. (C) Bar graphs show average (n = 50 per condition) ciliary length, polycystin-2 per unit length, and polycystin-2 per cilium. To obtain polycystin-2 per unit length, the amount of polycystin-2 in each cilium was divided by its length. Error bars, SEM.