(A–C) Decrease in mRNA and protein expression of IL-6 and TNF induced by Itgb1, Rps3, or Ywhaz. Mouse splenocytes (A) and peritoneal macrophages (B and C) were cultured with multiple concentrations (0, 10, 50, or 100 ng/mL) of recombinant Itgb1, Rps3, or Ywhaz in the absence or presence of 10 ng/mL LPS for 6 hours (for peritoneal macrophages) or 24 hours (for splenocytes). IL-6 and TNF protein concentrations in the culture supernatants were measured by ELISA (A and B), and their mRNA levels in the cells were determined by qRT-PCR (C). (D) Increase in IL-10 mRNA (top) and protein (bottom) expression induced by the 3 regulators. (E) Increase in Il6 and Tnf expression induced by the knockdown of Itgb1, Rps3, or Ywhaz transcripts. Peritoneal macrophages were transfected with siRNAs for Itgb1, Rps3, or Ywhaz for 24 hours. (F) Promotion of Il6 and Tnf production by anti-Rps3 or anti-Ywhaz Abs for 6 or 12 hours. (G) Decrease in IL-6 mRNA (top) and protein (bottom) expression in mouse fibroblast-like synoviocytes (FLSs) after treatment with recombinant Itgb1, Rps3, or Ywhaz for 6 hours. (H) Increase in mRNA levels of Il6 and Il8 in FLSs transfected with Itgb1, Rps3, or Ywhaz siRNAs. For qRT-PCR assays (C–H), mRNA levels of the target gene were first normalized to those of Gapdh, and then further normalized to the mean mRNA expression levels in the cells treated with LPS only (C, D, and G), those without treatment (none; F), or those transfected with control siRNA (E and H). Data are the mean ± SEM of more than 2 (F) or 3 independent experiments (A–E, G, and H). *P < 0.05, **P < 0.01, ***P < 0.001 by 1-way ANOVA with a post hoc test (Dunnett’s correction; A–D, F, and G) or Student’s t test (E and H).