title: GSM2858342: AT2_P60; Mus musculus; RNA-Seq
library_construction_protocol: Sftpc-CreER; Rosa26-Zsgreen; Hopx-RFP mice at various developmental stages were injected with 1-2 doses of tamoxifen. For AT1 cell isolation, lungs were inflated with an enzyme cocktail solution containing neutral protease (5U/ml,LS02111, Worthington), Elastase (4 U/ml, LS002294, Worthington), Type I collagenase (200 U/ml, 17100017, Gibco), and DNase I (0.33 U/ml, 10104159001, Roche) and incubated with the enzyme cocktail solution at room temperature for 45 minutes. Lungs were then cut into small pieces and filtered through 40μm cell strainers (BD Falcon). Cells were first incubated with red blood lysis buffer. After spinning down, cells were resuspended in DPBS+5% FBS (Gibco, 1767839). Cells were then stained with different antibodies. Antibodies used: PE-Cy™7 rat anti-CD45 (1:400, 552848, BD biosciences) and APC hamster anti-Pdpn (1:200, 127410, biolegend). For AT2 cell isolation, lungs were inflated with enzyme cocktail solution containing neutral protease (5U/ml, LS02111, Worthington) and DNase I (0.33 U/ml, 10104159001, Roche). DAPI was used to label dead cells. Sorting was performed on BD Aria2 RNA libraries were prepared for sequencing using 10X genomics single cell 3' standard protocols
attributes.tag: GEO Accession
attributes.value: GSM2858342