Skip to main content
. Author manuscript; available in PMC: 2011 Dec 24.
Published in final edited form as: Cell. 2011 Jun 24;145(7):1129–1141. doi: 10.1016/j.cell.2011.05.030

Figure 6. Elimination of primary cilia in vivo diminishes Notch signaling and inhibits basal to spinous cell fate commitment in developing epidermis.

Figure 6

(A) Confocal images showing that Notch reporter activity (GFP) is induced suprabasally in E17.5 spinous epidermal cells expressing scrambled but not Ift74 shRNAs. Transduced cells are H2B-RFP+. Quantifications reveal 5X less GFP reporter expression in Ift74-deficient vs. control cells. Histogram represents averaged data from 2–3 embryos in which >200 RFP+ cells were analyzed. *indicates p<.001 by students t-test. Error bars are +/− SEM. Scale bar=10µM. (B) H2B-RFP(+) Ift74-shRNA transduced areas show reduced K10 compared to adjacent uninfected or scrambled shRNA-transduced epidermis. Kif3a cKO epidermis similarly shows diminished K10 in comparison to WT. (C) Transgenic expression of NICD-GFP rescues Ift74 shRNA-induced defects in K10 and Hes1 expression. (D–E) Repression of differentiation markers Hes1, filaggrin and invoIucrin correlates with loss of cilia rather than proliferative status. Transduced areas of hyperproliferative Ift74-shRNA-knockdown epidermis (D) (H2B-RFP+) show reduced differentiation compared to scrambled controls. *denotes background fluorescence; arrows note nuclear Hes1. Hypoproliferative Kif3a cKO epidermis (E) shows similar diminished differentiation. (F) Failure to exclude blue dye indicates defective skin barrier formation in Ift74-shRNA- transduced and Kif3a cKO E17.5 embryos. B–E Scale bars=45 µM except for Kif3a data, where bar=100 µM. See also Figure S5–6.