GSM8012881
Whole organs were then dissected in the following order: small intestine, large intestine, spleen, pancreas, lymph nodes (inguinal lymph nodes, abdominal aortic lymph nodes, axillary lymph nodes, cervical lymph nodes), stomach, thyroid gland, brain, heart, lung, thymus, kidney, adrenal gland, ovary, uterus, bladder, liver, muscle, bone marrow and spinal cord. Except for bone marrow, the samples were quickly transferred into cold DPBS, washed and then manually pulverized using a mortar and pestle within a liquid nitrogen bath. The pulverized tissues were suspended in 1mL of Nuclear Isolation Buffer (NIB) and followed by incubation for 1.5-3 min to isolate the nuclei. The bone marrow cell suspension was centrifuged at 350 × g for 5 min, and the pellet was directly resuspended in 1 mL of NIB for 2min incubation. The suspension of each tissue was transferred to 15 mL centrifuge tubes pre-filled with 5mL of WB (wash buffer, 0.2% BSA, 3 mM MgCl2 in DPBS), filtered through 40 μm strainers and centrifuged at 650 × g for 5 min at 4°C. We resuspended the nuclei in 1 mL of WB and filtered the suspension through 20 μm size strainers, pelleted at 650 × g for 5 min and resuspended in the appropriate amount of DPBS to give a final concentration of 25,000 nuclei per microlitre. Library preparations were performed with the UUATAC-seq protocol