Fig 2. Targeting PTK6 expression improves TNFα/IFNγ mediated barrier dysfunction.
A) YAMC were electroporated with PTK6 siRNA or vehicle control (TE) then plated on ECIS arrays and allowed to express for 48 hours. Monolayers were then treated with vehicle control or indicated cytokines then resistance measurements were recorded every 90 seconds B) Either WT or PTK6-/- YAMCs were grown on gold-plated ECIS arrays, stimulated with either vehicle control (VC) or TNFα (100ng/ml) and IFNγ (500U/ml), then resistance measurements were recorded every 90 seconds. Values were normalized to timepoint zero and shaded area represents standard error (n = 4). C) PTK6-/- cells were transfected with empty vector (EV) or PTK6 cDNA then treated with either vehicle control or TNFα (100ng/ml) and IFNγ (500U/ml). Inlay shows successful overexpression of PTK6 cDNA vs. EV transfected cells. D) Either WT or PTK6-/- YAMCs were grown on transwell inserts at 2 X 105 cells/ml until reaching 2-days post-confluence then treated with vehicle control (VC) or TNFα (100ng/ml) and IFNγ (500U/ml) for 24 hours. Permeability coefficients were calculated as described in methods. Error bars represent standard error (*p<0.05 compared to WT, n = 3). E) Westerns showing relative expression of PTK6 in siRNA treated and epithelial cells from PTK6-/- knockout mice (Please see S4 Fig for full blot scans).