Fig. 10

ARPE-19 cells under severe stress conditions like the traumatic destruction of the cell monolayer by a scratch 24 h prior staining for FoxP3. A The cell layer of confluently grown ARPE-19 cells was scratched (here: not treated with IL-1β) 24 h prior to staining for FoxP3. The scratched cells showed a translocation of FoxP3 from the cytoplasm to the nucleus (left panel: FoxP3 predominantly localized perinuclear and nuclear), while in confluent, unstressed cells FoxP3 expression is seen in the cytoplasm (right panel: confluent, untreated). Scale bar represents 20 µm. B Secretion of VEGF-A, MCP-1, IL-8 and IL-6 (pg/ml) by confluent ARPE-19 where the cell monolayer was scratched 24 h prior to the additional stimulation with IL-1β for 6 min, 1 and 2 h. (IL-6 = interleukin-6; IL-8 = interleukin-8; MCP-1 = monocyte chemoattractant protein-1 (CCL2); VEGF-A = vascular endothelial growth factor-A). Triplicate cultures of ARPE-19 were treated as described; culture supernatants were collected at the end points of IL-1β-treatment. Supernatants were pooled and tested for cytokine and chemokine secretion as duplicates in a multiplex bead assay (N = 2). The final values of duplicate supernatants are calculated by the standard curves of the assay from the median fluorescence intensity of at least 50 beads measured for each analyte and sample