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. 2011 Jun 20;108(27):11163–11168. doi: 10.1073/pnas.1107899108

Fig. 5.

Fig. 5.

ZLD reduces IRAK-M in human PBMCs and enhances LPS induced responses. (A) Human PBMC from 11 independent donors were pretreated (first treatment) with 10 μM ZLD and then 100 pg/mL LPS for an additional 24 h. The supernatants were tested for IL-8 by ELISA. The values were normalized by the value of cells treated with vehicle and then treated with LPS (9.19 ± 2.08 ng/mL). Nonresponders are shown in black and dashed line represents 100%. (B) Triplicate wells of PBMC from a single responsive donor were pretreated with vehicle, 10 μM ALD, or 10 μM ZLD for 24 h and then treated with vehicle or 1 ng/mL LPS for 24 h. IL-8 in the supernatant was measured by ELISA. *P < 0.05 by one-way ANOVA with Dunnett's post hoc test. (C) Responder PBMCs (donors 1 and 2) and nonresponder PBMCs (donors 3 and 4) were treated with 10 μM ZLD or vehicle for 24 h. The lysates were immunoblotted and probed for IRAK-M, IRAK-1, and β-actin, as shown. (D) PBMCs from a representative responsive donor were pretreated with vehicle (Veh), 10 μM ZLD, or 10 μM ZLD and 80 μg/mL farnesyl pyrophosphate (ZLD+FPP) for 24 h, then treated with 1 ng/mL LPS or vehicle for 24 h in triplicate. *P < 0.05 by one-way ANOVA with Dunnetts post hoc test.