Fig. 3
Validation of the gating strategy using AT2 cells with intrinsic GFP expression. a Scgb1a1-rtTA/(tetO)7CMV-Cre mice were bred with ROSAmTmG mice to generate triple-transgenic mice that express EGFP in AT2 cells following cre-mediated recombination. b Immunofluorescence analysis of the lung from recombinant Scgb1a1-rtTA/(tetO)7CMV-Cre-mTmG mice demonstrated that proSP-C+ AT2 cells express GFP. AT1 cells do not express GFP. Scale bars, 50 μm. c GFP+ cells of single lung cells (left) from recombinant Scgb1a1 (Line 1)-rtTA/(tetO)7CMV-Cre-mTmG mice were analyzed in the dot plot of EpCAM and MHCII (right). Almost all GFP+ cells were gated with P1 cells. d FACS analysis of sorted GFP+ cells demonstrates that sorted cells are primarily positive for proSP-C (right) compared to controls (left). EGFP, enhanced green fluorescent protein; tdTomato, tandem-dimer Tomato; EpCAM, epithelial cell adhesion molecule; MHCII, major histocompatibility complex II; proSP-C, pro-surfactant protein C; SSC, side scatter; DAPI, 4′,6-diamidino-2-phenylindole