Fig. 6

The direct actions of JAKis and the TNFi on the regulatory machinery of rheumatoid arthritis (RA) susceptibility genes. a, e Organization of transcriptional regulatory regions around the TRAF1 (a) and CD40 (e) gene. The boxed area indicates a putative enhancer that overlaps with RA risk SNPs. SS (synergistic stimuli) means a combination of eight different cytokines (IFN-α, IFN-γ, TNF-α, IL-1β, IL-6/sIL-6R, IL-17, TGF-β1, and IL-18) [26]. Data were visualized using the Integrative Genomics Viewer (IGV). b, f A schematic image of insertions into luciferase reporter vectors. c, g Relative luciferase activity of an allele-specific reporter assay (n = 3) using HT-1080 cells. Bars, mean; error bars, SD. P values were determined using paired t-test (*P < 0.05). d, h Transcript abundances of TRAF1 (d) and CD40 (h) obtained from qRT-PCR data in genetically edited cells (n = 3). A small region surrounding rs7021049 or rs6074022 was deleted in MH7A cells using the CRISPR/Cas9 system. Bars, mean; error bars, SD. P values were determined using paired t-test (*P < 0.05). i A graphical summary of the effector sites of JAKis and the TNFi on the CD40-TRAF1 cascade. ABC activity-by-contact, ADA adalimumab, BARI baricitinib, NS non-stimulated, NT non-treated, SS synergistic stimuli, TOFA tofacitinib, UPA upadacitinib, 24 h 24 h, 7d 7 days