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. 2021 Feb 15;10:e59119. doi: 10.7554/eLife.59119

Figure 7. BBS3 is essential for PLD to associate with the BBSome for ciliary exit.

Figure 7.

(A and B) Immunoblots of whole-cell extracts (WCE) (A) and ciliary extracts (CE) (B) of CC-5325 and clip1 cells probed with α-BBS3 and α-PLD. (C) CC-5325 and clip1 cells stained with α-PLD (red). (D and E) Immunoblots of WCE (D) and CE (E) of CC-125, BBS3miRNA, BBS3Res-WT, BBS3Res-A73L, and BBS3Res-△N cells probed with α-BBS3, α-BBS1, α-BBS5, and α-PLD. (F) CC-125, BBS3miRNA, BBS3Res-WT, BBS3Res-A73L, and BBS3Res-△N cells stained with α-PLD (red). (G) Immunoblots of sucrose density gradient of CE of CC-125 and BBS3Res-△N cells probed with α-BBS1 and α-BBS5. (H) Immunoblots of sucrose density gradient of CE of BBS3Res-A73L, BBS3Res-△N, and clip1 cells probed with α-BBS1, α-BBS5, and α-PLD. (I) Schematic presentation of how BBS3 binds and recruits the BBSome to the ciliary membrane for interacting with PLD and how PLD-associated BBSome loads onto IFT trains for ciliary exit. For panels A, B, D, and E, α-tubulin and Ac-tubulin were used to adjust the loading for WCE and CE, respectively. For panels C and F, the inset shows ciliary tips. Scale bar 10 µm.