Fitc anti mouse cd326 epcam

Frozen sections of lacrimal gland tissue were fixed with methanol/acetone (1:1), blocked using 5% normal goat serum (WAKO)/0.3% Triton×-100 (Sigma) in phosphate-buffered saline (PBS), and stained with FITC anti-mouse CD326 (EpCAM) (118207, BioLegend), AdipoR2 (sc-514045), and PPARγ (2443) antibodies. Alexa Fluor594-conjugated anti-mouse IgG (H+L) (8890, Cell Signaling Technology) and Alexa Fluor 555-conjugated anti-Rabbit IgG (H+L) (A21428, Thermo Fisher Scientific) were used as secondary antibodies. These antibodies were diluted with Can Get Signal immunostain solution (Toyobo). After washing 3 times with PBS, nuclear DNA was stained with ProLong Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific). Sections were observed using a fluorescence microscope (KEYENCE) at a magnification of 400× or 1000×.

Adult murine intestinal organoids in Matrigel/collagen type I and dFEnS were dissociated into single cells by incubating fragmented organoids in 50% v/v TryPLE Express (Gibco, #12605-101) in advanced DMEM/F12 for 10–15 min. These single cells were stained with FITC anti-mouse CD326 (Ep-CAM) (Clone G8.8, BioLegend, 1:500, #118207), PE anti-mouse Ly6A/E (Sca-1) (Clone D7, BioLegend, 1:5000, #108107), and propidium iodide (Immunostep, 1:1000, #PI) before flow cytometry analysis with FACS Melody (BD Biosciences, Franklin Lakes, N). Live single cells were gated to assess for Sca-1 and Ep-CAM.