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Fig. 2. MYB is expressed early and transiently during multiciliogenesis in the lung and brain. (A,B) Immunostaining of E17.5 mouse lung sections for MYB and either SCGB1A1 (Clara cells) or FOXJ1 (MCCs). MYB is not detected in SCGB1A1-expressing cells (boxed region, insets in A) but many MYB-positive cells also express FOXJ1 (boxed region, insets in B). (C) Sections of E14.5 through E17.5 lungs were stained for MYB, FOXJ1 and SOX2 and the number of epithelial cells (marked by SOX2) expressing MYB only, FOXJ1 only, or both MYB and FOXJ1 were counted. Total SOX2-positive cells scored was >1000 per age. (D) Mouse tracheal epithelial cell (MTEC) cultures were immunostained for MYB and FOXJ1 at the times indicated after exposure to an air-liquid interface (ALI) to promote differentiation. The number of cells that were MYB+, FOXJ1+ or double-positive were counted. Similar to in vivo, the FOXJ1+ population continually increased during culturing, whereas the percentage of MYB+ and double-positive cells initially increased and then declined. Total cells scored was >100 per time point. (E) Immunostaining of E17.5 lung section for MYB, FOXJ1 and acetylated tubulin (ACT, cilia marker). MYB is seen in developing (upper inset) but not mature (lower inset) MCCs. (F) Immunostaining of ALI +8 days MTEC cultures for MYB and ACT. MYB is not detected in mature MCCs. Arrowheads (E,F), ciliary tufts of mature MCCs. (G,H) Whole-mount ventricles of P2 mouse brain stained for FOXJ1 and MYB (G) or Ki67 and MYB (H). (I) Stage-specific expression of MYB during multiciliogenesis in MTECs. ALI +6 days MTECs were stained for MYB and pericentrin (PCNT), a centriole marker, and cells were staged as described in Fig. 1A. MYB is detected at stages I and II, before and during centriole amplification, but not during stages III or IV. Dotted lines show cell outline. Scale bars: 10 μm in A,B,E-H; 5 μm in I. Image published in: Tan FE et al. (2013) © 2013. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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