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Tissue collection and microscopic analysis Spleens were fixed in 4% formaldehyde (methanol-free paraformaldehyde, Thermo Scientific #28908) overnight at 4 ยฐC, washed and transferred into phosphate-buffered saline containing 0.02% sodium azide and 20% sucrose and then cut with a microtome (CryoStar NX 50, Thermo Scientific) to obtain 8 ยตm tissue sections 94 that were then stained for 10 min with 0.3 ยตM 4โฒ,6-diamidino-2-phenylindole in Dulbeccoโs phosphate-buffered saline (Thermo Scientific #62247)
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