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. 2023 Jun 5;24(7):1200–1210. doi: 10.1038/s41590-023-01527-9

Extended Data Fig. 5. Evidence of STAT1 and AP-1 activation in FLS.

Extended Data Fig. 5

a, Percentage of cells staining for nuclear pSTAT1 or cJun found within the synovial lining, which was manually annotated based on tissue architecture, in confocal images from Fig. 4d (n = 3 tissues, 6 sections). Error bars: mean ± standard deviation. b, Representative confocal images of a lymphocyte aggregate within the sublining stained for phosphorylated STAT1 (cyan), PDPN (red), CD3 (magenta) and nuclear marker (blue) (n = 4 tissues, 8 sections). White arrows indicate FLS with nuclear pSTAT1. c, Representative confocal images of a lymphocyte aggregate stained for the antigens in (b) above as well as the colocalization of HLA-DR and PDPN (yellow) as defined by pixels with fluorescence intensity in the top 10% for both HLA-DR and PDPN (n = 3 tissues). d, Percent of genes identified in the multiome dataset with accessible AP-1 motifs in their promoters that were up- or down-regulated in cultured FLS stimulated with the indicated cytokines. AP-1 target genes (1024 total) in the activated lining FLS state were identified as those with motif matches with a log-odds score of at least 10 (computed by motifmatchr). e, Dynamics of chromatin accessibility of the STAT1 motif containing cis-regulatory elements across ex vivo FLS states and in vitro cytokine stimulated or unstimulated FLS. Empirical cumulative distribution function (ECDF) 600 plots of ChromVAR z-scores for STAT1 motif from scATAC-seq analysis of the indicated cultured FLS samples or FLS states are shown.

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