Human decidual tissues were briefly fixed in 4% paraformaldehyde (PFA) and embedded in O.C.T. Compound (Catalog No. 4583, Sakura Finetek, Torrance, CA). The frozen sections at 10 μm were further fixed in 4% PFA and treated with 0.1% triton, and subjected to the incubation with specific antibodies against NCAM1/CD56 (Catalog No. ab75813, Abcam, Cambridge, MA), CK7 (Catalog No. ab181598, Abcam), CD39 (Catalog No. 14211-1-AP, Proteintech, Wuhan, China), or CD103 (Catalog No. 350227, BioLegend). Binding of the antibody was visualized using FITC-conjugated or TRITC-conjugated secondary antibody (Catalog No. ZF-0311 or ZF-0313, ZSGB-BIO, Beijing, China), and cell nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI; Catalog No. 28718-90-3, Sigma). Immunofluorescent staining was examined using Zeiss LSM780 confocal system (Carl Zeiss, Jena, Germany) and processed with ZEN 2012 software (Carl Zeiss). Immunohistochemical staining for CK in decidua was performed by using antibody against CK7 (Catalog No. ab181598, Abcam) and HRP-conjugated second antibody (Catalog No. PV-6001, ZSGB-BIO) followed by recovery of substrate diaminodbenzidine (DAB) (Catalog No. ZLI-9019, ZSGB-BIO). The imagines were recorded on a light microscope with charge-coupled device (CCD) (Olympus, Tokyo, Japan).
Wang F., Jia W., Fan M., Shao X., Li Z., Liu Y., Ma Y., Li Y.X., Li R., Tu Q, & Wang Y.L. (2021). Single-cell Immune Landscape of Human Recurrent Miscarriage. Genomics, Proteomics & Bioinformatics, 19(2), 208-222.
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