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. 2014 Mar 1;25(5):643–657. doi: 10.1091/mbc.E13-07-0429

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© 2014 Rozycki et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — Prolonged increase in myosin activity is sufficient and necessary for cilium loss. (A) Confluent LLC-PK1 cells were transfected with the constitutively active Myc-tagged (DD) MLC II in serum-free medium and stained 24 h later for Myc and Ac-tub. (B) The presence of the cilium was quantified in DD-MLC–expressing cells and their nontransfected neighbors on the same coverslip (mean ± SEM, n = 3, 25 cells/condition). (C) LLC-PK1 cells were transfected with nonrelated (NR) or myosin heavy chain II (MHC II)–specific siRNA, grown to confluence, treated as indicated for 48 h, and then stained for Ac-tub. MHC II knockdown was verified by Western blotting. (D) Ciliation percentage was quantified (mean ± SEM, n = 3, ∼150 cells/experiment). (E) LLC-PK1 cells were transfected with siRNAs against MRTF A and MRTF B or NR siRNA. After reaching confluence, cells were treated as shown for 48 h and then subjected to Western blotting for the indicated proteins. (F) LLC-PK1 cells treated as in E were stained for Ac-Tub. (G) Ciliation percentage for E was quantified (mean ± SEM, n = 3, ≈150 cells/experiment).