Lsp 880

Stomach tissues were harvested and fixed in 4% paraformaldehyde (PFA) before paraffin embedding. H&E staining, immunohistochemical staining (IHC), and immunofluorescence staining (IF) were carried out on 5 µm paraffin-embedded sections. Antibodies were as follows: GFP (1:200, Cat. no. 2956, CST, Danvers, MS), BrdU (1:300, Cat. no. ab6326, Abcam, Cambridge, UK), Ki67 (1:1000, Cat. no. ab15580, Abcam), E-cadherin (1:300, Cat. no. 610181, BD Biosciences, Franklin Lakes, NJ), cleaved caspase-3 (1:200, Cat. no. 9661S, CST), TRITC-labelled UEA (1:100; Cat. no. L4889, Sigma), ChrgA (1:100, Cat. no. ZA-0066, Zhongshanjinqiao, Beijing, China), Tff2 (1:500, R33452, Sigma), FITC-labelled DBA (1:100, L9142, Sigma), RFP (1:500, Cat. no. 600-401-379, Rockland, Limerick, PA; tdTomato can be recognized by the RFP antibody), p53 (1:100, Cat. no. 2524S, CST,). The immunofluorescence staining was visualized with TSA regents (Fluorescein, FP1168; Cyanine3, FP1170; Cyanine5, FP1171; PerkinElmer, Waltham, MA) and nuclear counterstaining was performed using DAPI. IF was observed under confocal microscopy (LSP 880, Zeiss, Jena, Germany). IHC was visualized with DAB (1:20, Cat. no. ZLI-9018, Zhongshanjinqiao).

Harvested gastric antral tissues were fixed in 4% PFA for 1 hour. For the whole mount analysis of endogenous GFP, gastric tissues removed from the muscle layer were mounted upside down. For the frozen section, tissues were embedded in O.C.T. (4583, Sakura tissue-Tek, Tokyo, Japan) and then cut into sections using a Cryostat microtome (77200186 Issue 5, Thermo, Waltham, MA). Nuclear counterstaining was performed using DAPI and endogenous GFP was directly imaged under confocal microscopy (LSP 880, Zeiss).