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. 2021 Sep 14;12:715997. doi: 10.3389/fimmu.2021.715997

Figure 2.

Figure 2

FLSs internalize carbamylated LL37 and induce antigen-specific autoantibody responses. NETs containing carbamylated LL37 were incubated with RA FLS for 24 hours. (A) Intracellular co-localization of carbamylated LL37 and MHCII was assessed in RA FLS. Red, LL37; green, MHCII; blue, DNA. (B) Co-localization of MHCII and LL37 was assessed in the plasma membrane of un permeabilized RA FLS after 48h incubation with NETs. Green, LL37; red, MHCII; blue, plasma membrane. Results are representative of three independent experiments. Original magnification is 400X. (C) Serum samples from HLA-DRB1*04:01 transgenic mice immunized with FLS alone or FLS that internalized RA-NETs were analyzed for the presence of autoantibodies against carbamylated LL37. Results are the mean +/- SEM of n=3-5 per group. One-way ANOVA- Dunn’s multiple comparisons test was used. *p < 0.05. (D) Autoantibodies against carbamylated LL37 were measured in non-RA (n = 9) and RA (n = 15) synovial fluid. Black line = 2 standard deviations above mean for non-RA patients. Results are the mean +/- SEM. Mann-Whitney U test was used. **p < 0.01.