Article Title: A single cell atlas of the tracheal epithelium reveals the CFTR-rich pulmonary ionocyte
Figure Lengend Snippet: Pulmonary ionocytes are a major source of CFTR activity. a , Immunofluorescence for Krt5 (green) and Foxi1 (red) in mouse tracheae at homeostasis (left, n=3) or 3 days post injury (right, n=3). Arrowhead: Foxi1 + Krt5 + cells. Arrow: Foxi1 + Krt5 - cell. Scale bars, 20μm. b , Immunofluorescence and quantification for ionocytes (FOXI1 + , red, arrowheads) and ciliated cells (FOXJ1 + , green) in HBEC cultures treated with DMSO or DAPT, scale bar, 100 μm, (n=4 experiments in one donor). *p-value=0.01, ***p-value=1.1×10 −6 by two-tailed t-test c , HBECs were treated with DMSO or DAPT upon culture at ALI. After differentiation (2–3 weeks), cultures were loaded into Ussing chambers and short circuit current (I sc ) was recorded during addition of Amiloride, Forskolin, and a CFTR-inhibitor, CFTR(inh)-172. Shown is a representative tracing from Donor 1 (n=11). d , Change in short circuit current (ΔI sc ) in response to Forskolin measured in DMSO (n=7 cultures per donor) and DAPT-treated cultures, (n=8 cultures per donor). ***p-value
Article Snippet: Amiloride (Sigma, A9561) was added apically at 10μM to inhibit Na+ absorption, then Forskolin (Sigma, F6886) was added apically at 20μM to stimulate cAMP and finally, CFTR-172 (Sigma-Aldrich, C2992) inhibitor was added apically and basally at 30μM.
Techniques: Activity Assay, Immunofluorescence, Two Tailed Test