Fitc conjugated anti mouse cd326 epcam mab

Frozen sections of salivary gland tissue were fixed with methanol/acetone (1:1), blocked using 5% normal goat serum (WAKO)/0.3% Triton X-100 (Sigma) in phosphate-buffered saline (PBS), and stained with FITC-conjugated anti-mouse CD326 (EpCAM) mAb (118207, BioLegend), Alexa Fluor 647-conjugated anti-mouse CD4 mAb (100426, BioLegend), anti-mouse CD153 mAb (14-1531-85, Thermo Fisher Scientific), and an anti-mouse CXCL13 polyclonal antibody (PA1-29046, Thermo Fisher Scientific). Alexa Fluor 555-conjugated anti-rat IgG (H+L) (4417) and Alexa Fluor 594-conjugated anti-rabbit IgG (H+L) (8889), which were obtained from Cell Signaling Technology, were used as the secondary antibody. These antibodies were diluted with the Can Get Signal immunostain solution (Toyobo, Osaka, Japan). After washing 3 times with PBS, nuclear DNA was stained with ProLong Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific). Images were acquired using the KEYENCE BZ-X800 microscope (Osaka, Japan), and processed with the full focus function of the BZ-X800 Analyzer software (Itasca, IL, USA)unless otherwise stated.

Immune cells and epithelial cells harvested from tissues were stained using a PE-Cy7-conjugated anti-mouse CD4 monoclonal antibody (mAb) (100405), PE-Cy5-conjugated anti-mouse CD8a mAb (100709), PE-Cy5-conjugated anti-mouse/human CD11b mAb (101209), allophycocyanin (APC)-Cy7-conjugated anti-mouse CD11c mAb (17323), APC-conjugated anti-mouse F4/80 mAb (123115), PE-conjugated anti-mouse CD19 mAb (152407), PE-Cy7-conjugated anti-mouse CD45.2 (Ly5.2) mAb (109829), APC-conjugated anti-mouse CD44 mAb (103011), APC-Cy7-conjugated anti-mouse CD62L mAb (104427), PE-conjugated anti-mouse CD153 mAb (106405), PerCP/Cy5.5-conjugated anti-mouse CD184 (CXCR4) mAb (146509), PerCP/Cy5.5-conjugated anti-mouse CD185 (CXCR5) mAb (145507), fluorescein isothiocyanate (FITC)–conjugated anti-mouse CD279 (PD-1) mAb (135213), and FITC–conjugated anti-mouse CD326 (EpCAM) mAb (118207), which were obtained from BioLegend (San Diego, CA, USA). A Canto II flow cytometer (BD Biosciences, San Jose, CA, USA) was used to identify cell populations according to surface expression profiles. Flow cytometric data were analyzed using FlowJo software v10 (BD Biosciences). The representative gating strategy for analyzing SA-T cells was reported previously [51 (link)].