Figure 3.

Expression and production of Th17-inducing cytokines by human limbal epithelial cells (HLECs) exposed to TNF-α ([A] and [B]) and hyperosmotic media ([C] and [D]). The HLECs were treated with TNF-α (10 ng/ml) or hyperosmotic media (450 mOsM) for 4–48 hours in the absence or presence of 5 µM NF-κB-I or 10 ng/ml GDNF, which was preloaded to the medium for 1 hour. The Th17-inducing cytokines (TGF-β1, IL-6, IL-23, and IL-1β) were measured by reverse transcription and quantitative real-time polymerase chain reaction for mRNA ([A], [C]) and by enzyme-linked immunosorbent assay and Luminex immunobead assays in culture supernatants ([B], [D]). Results shown are mean ± SD of four independent experiments. *, p < .05; **, p < .01; by t test. Abbreviations: GDNF, glial cell-derived neurotrophic factor; IL, interleukin; NF-κB-I, NF-κB inhibitor; TGF, transforming growth factor; TNF-α, tumor necrosis factor-α.