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. 2010 Sep 27;107(41):17692–17697. doi: 10.1073/pnas.1009443107

Fig. 7.

Fig. 7.

GSK3α/β controls GA-induced sIL-1Ra production. (A) Monocytes were stimulated by GA for indicated time, and cell lysates were subjected to Western blot analysis. (B and C) Monocytes were preincubated for 45 min in the presence or absence of 10 μM SB216763 (SB) and then cultured in the absence (white columns) or presence (gray columns) of GA. (B) Production of sIL-1Ra was measured in harvested supernatants (no inh. = 100% = 2020 ± 636 pg/mL sIL-1Ra) and (C) mRNA analyzed by real-time quantitative PCR; results are presented as described in Fig. 2 B and D, respectively. (D) Monocytes were nucleofected with stealth siRNA for GSK3α and GSK3β (siGSK3) or negative control (mock). Efficiency of GSK3α/β silencing was assessed by Western blot (Lower). sIL-1Ra was measured in culture supernatants of GSK3α/β knocked-down or mock-transfected monocytes activated (gray columns; 100% = 2174 ± 467 pg/mL sIL-1Ra) or not activated (white column; sIL-1Ra concentration = 1,344 ± 676 pg/mL) by GA. (E) Monocytes were preincubated in the absence or presence of 10 μM Ly294002 (Ly); 5 μM IC87114 (IC); 500 nM PI3Kγ inhibitor, 5 μM U0126 (U0), or 5 μM PD98059 (PD) before activation by GA. Cell lysates were subjected to Western blot analysis. **P < 0.01 as determined by Student's t test.