Anti epcam clone g8

Tissues were processed as previously described⁵¹ . Briefly, tissues were fixed in paraformaldehyde (PFA), lysine, and sodium periodate buffer (PLP, 0.05M phosphate buffer, 0.1M L-lysine, pH 7.4, 2 mg/mL NaIO₄, and 10mg/mL PFA) overnight at 4°C followed by 30% sucrose overnight at 4^oC and subsequent embedding in OCT media. Frozen tissues were sectioned at 20μm using Leica CM3050S cryostat, and FcR blocked with anti-CD16/32 Fc-block (Clone 93, BioLegend) diluted in PBS containing 2% donkey serum, 2% fetal bovine serum (FBS), and 0.1% Triton-X for 1h at 25^oC. Sections were stained with anti-CD8α (Clone 53–6.7, BD Bioscience), anti-EpCAM (clone G8.8, eBioscience), anti-Influenza A virus (polyclonal, Abcam) and anti-CD11c (clone N418, BioLegend) diluted in PBS containing 2% goat serum, 2% FBS, 0.1% Triton-X, and 0.05% Fc block for 1h at 25^oC. Images were acquired using a Zeiss LSM 880 confocal microscope (Carl Zeiss) with the Zen Black software. The imaging data were processed and analyzed using Imaris software version 9.0.1 (Bitplane, Oxford Instruments).