Luteolin inhibits Th17 cell differentiation and spinal infiltration in RA

Cut&tag was applied to explore the mechanism of NFATC2 confers Th17 cell differentiation and migration in CIA. Overall design: CD4+T cells from CIA and control mice were harvested, counted, and centrifuged at 600g for 3min at room temperature. A total of 50,000 cells were subject to the CUT&Tag assay performed by SeqHealth (Wuhan, China) using a commercial kit (Vazyme, China, TD903) according to the manufacturer's protocol. Briefly, cells were incubated with Concanavalin A-coated magnetic beads. Subsequently, anti-NFATC2 or IgG control antibody were added, and the cells were incubated on a nutator overnight at 4?. After that, secondary antibody was performed on a nutator for 1h at room temperature followed by incubation with hyperactive pG-Tn5 transposonase. After transposon activation and tagmentation, DNA was extracted, amplified, and purified to construct library. Sequencing was performed in the Novaseq 6000 sequencer (Illumina) with PE150 model.