Fig. 4

IMG cells respond to LPS treatment by increasing protein expression of inflammatory cytokines. a ELISA was used to analyze the secretion of TNF-α and IL-6 into IMG cell-conditioned medium during a 16-h incubation with or without LPS (10 ng/mL) or IL-4 (10 ng/mL) as described in the “Methods” section. b ELISA analysis of IL-1β in whole-cell lysates of IMG cells treated as described in (a). c ELISA of conditioned media (filled bars) and cell lysates (open bars) collected from IMG cells treated with or without LPS (10 ng/mL) for 6 h. LPS-treated cells were also incubated with or without Ac-YVAD-CMK (40 μM; YVAD) for 5 min, followed by ATP (5 mM) for 30 min as indicated. d Immunoblot analysis of conditioned media from IMG cells treated as described in (c). Blots were probed for IL-1β. Pro- (34 kDa) and mature (17 kDa) IL-1β bands are indicated. One-way ANOVA was used to determine the significance of the data. *P < 0.01; **P < 0.0001. Data are represented as means ± s.d. (n = 3, technical replicates). Each experiment was repeated on at least two separate occasions