Fig. 1
LPS enhances the EtdBr dye uptake in airway epithelial cells. a-b EtdBr dye uptake rates (fluorescence intensities (AU)/min) under different perfusion conditions (exemplary experiment see Fig. S1) in Calu-3 cells (a) and PBEPCs (b) cultivated under control conditions or with 1 ng/mL LPS for 24 h. Perfusion conditions: 2 mM [Ca2+]ex, 0 mM [Ca2+]ex or 0 mM [Ca2+]ex + 1 mM La3+ (n = cell patches for Calu-3 cells, single cells for PBPECs). Kruskal-Wallis test with Dunn’s multiple comparison test (p < 0.05 *, p < 0.001 *** vs. different perfusion conditions; p < 0.05 #, p < 0.001 ### vs. control). c Real-time qRT-PCR for different Cx isoforms in PBEPCs. d Immunofluorescence staining against Cx26 or Cx43 (yellow) in PBEPCs. Scale bar = 20 μm. e Exemplary immunofluorescence staining against Cx26 (yellow) in the airways of PCLS cultivated under control conditions or with 1 µg/mL LPS for 3 h. Scale bar = 10 μm. f Cx26 immunofluorescence signal (particle count/area, relative to control) in PCLS after treatment with 1 µg/mL LPS for 3 h (n = analyzed PCLS from 3 donors, 2 PCLS/donor with mean of 3 airway areas/PCLS). Different symbol shapes visualize different donors. Unpaired two-tailed Student’s t-test (p < 0.05 * vs. control). g EtdBr dye uptake rates in absence of [Ca2+]ex relative to the rates obtained in presence of [Ca2+]ex in Calu-3 cells in which Cx26 or Cx43 expression was suppressed using respective siRNA and which were treated with 1 ng/mL LPS or vehicle for 24 h (n = measured cell patches). Kruskal-Wallis test with Dunn’s multiple comparison test (p < 0.01 **, p < 0.001 *** vs. negative siRNA + vehicle; p < 0.05 #, p < 0.001 ### vs. respective siRNA + vehicle; p < 0.001 +++ vs. LPS). h EtdBr dye uptake rates in absence of [Ca2+]ex ± 5 µM CVB4-57 relative to the rates obtained in presence of [Ca2+]ex in Calu-3 cells treated with vehicle or 1 ng/mL LPS for 24 h (n = cell patches). Kruskal-Wallis test with Dunn’s multiple comparison test (p < 0.001 *** vs. vehicle; p < 0.001 ### vs. LPS)