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Figure 2. Localisation of CFAP161 and the dependence on FOXJ1. (A) Schematic representation of the exon structure of Cfap161 and regions encoding the peptides (pI and pII) used for the generation of antibodies. (B) Western blot of overexpressed (CFAP161Flag in CHO cells) and endogenous CFAP161 (from mouse testis lysate) detected with monoclonal (pI) and polyclonal (pII) antibodies. The full-size Western blots are shown in Fig. S9A. (C) Induction of CFAP161 expression during cilia formation of air–liquid interface (ALI) cultures of mouse tracheal epithelial cells (mTEC). α-CFAP161 pII was used for visualising CFAP161, α-IFT88 to monitor ciliogenesis and α-β-Tubulin (β-TUB) as loading control. The full-size Western blot is shown in Fig. S9B. (D) Detection of endogenous CFAP161 by indirect immunohistochemistry of E17.5 wild type and Foxj1-mutant sections. Red boxed areas in a-h indicate the regions shown at higher magnification in a′–h′. Note that CFAP161 is absent in all analysed tissues of Foxj1lacZ/lacZ specimens. α-CFAP161 pI was used for the indirect DAB-immunostaining. (E) Expression of cfap161 was largely erased in foxj1-crispant embryos (b, b′) in comparison to wild types (a, a′). Red boxed areas in (a, b) indicate the regions shown at higher magnification in (a′, b′). (F) Unilateral foxj1 gain of function (foxj1-GOF) induced cfap161. Side of injection as indicated by asterisk. Scale bars: (D) = 500 µm; (E, F) = 100 µm. |
