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Immunostaining was performed as previously described. For immunostaining, the gut sections were fixed, blocked, and incubated with primary antibodies against CCR7 (BioLegend, 1:200), CXCR5 (BioLegend, 1:200), and PD-1 (BioLegend, 1:200) at 4°C overnight. After washing, the sections were co-stained with 2 μg/mL 4′,6-diamidino-2-phenylindole (nucleus) for 5 min. Fluorescence images were acquired using a confocal microscope (Leica SP8) with a 20× objective.
Copyright and License information: ©2022 Cheng, Zhou, Li, Shen, Zhao, Liu, Zhong, Chang, Kermode and Qiu
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