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. 2020 Jan 1;16(2):216–227. doi: 10.7150/ijbs.38722

Figure 4.

Figure 4

sCD83 binds CD154 on the surface of sTh2 cells. (A) PBMCs were isolated from blood samples obtained from AR patients (sensitized to HDM) and cultured in HDM (5 µg/ml) overnight, and then exposed to sCD83 (labeled with FITC) in the culture for 6 h. The cells were stained with anti-CD4 and IL-4 antibodies, and analyzed by flow cytometry. Gated dot plots show the sCD83-labeled cells (indicate that the sCD83 bound to a portion of PBMCs); these cells were also CD4+ IL-4+ (B) and CD154+ (C). (D) CD83+ CD154+ cells were isolated by FCCS and analyzed by co-IP. Immunoblots show a complex of CD83/CD154. (E-F) Bars show immunoblot image analysis data of panel D. (G) Immunoblots show results of CD154-depletion in sTh2 cells. (H) bars show immunoblot image analysis data of panel G. (I) Bars show Bcl2L12 mRNA levels in sTh2 cells. Data of bars are presented as mean ± SEM. Each dot in bars presents data obtained from each experiment. CD154d: CD154-deficient sTh2 cells (prepared by CD154 CRSPR). Control: sTh2 cells were treated with control CRISPR reagents. IL-5: IL-5 in the culture (20 ng/ml). Statistics of H-I: ANOVA (p<0.0001) followed by Bonferroni test. Data represent 6 independent experiments.