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. 2018 Jun 29;9:1503. doi: 10.3389/fimmu.2018.01503

Figure 7.

Figure 7

Regulation of serum amyloid A (SAA)3-induced IL-22 expression in neutrophils. (A) Bone marrow neutrophils from WT or Tlr2−/− mice (both male and female, 8–10 weeks of age) were stimulated with recombinant SAA3 (rSAA3) (1 µM) or vehicle for 24 h. IL-22 release was measured by ELISA. (B,C) SAA3-induced phosphorylation of selected signaling molecules. Bone marrow neutrophils were treated with rSAA3 (1 µM) as indicated. Cells were collected at indicated time points and cell lysates were separated on SDS-PAGE for Western blotting using antibodies to the phosphorylated and total proteins of selected signaling molecules (n = 3–5 mice for each group). (D) Pharmacological inhibition of SAA3-induced IL-22 secretion. Bone marrow neutrophils were pre-treated with the ERK1/2 inhibitor PD98059 (100 µM), p38 inhibitor SB202190 (100 µM), JNK inhibitor SP600125 (10 µM), Akt inhibitor MK-2206 2HCl (10 µM), STAT3 inhibitor static (10 µM), and NF-κB inhibitor celastrol (100 µM) for 1 h before rSAA3 (or medium only) stimulation for 24 h. The released IL-22 was quantified by ELISA (n = 3–5 mice for each group).