Analysis of control and stable Verge-expressing RBE4 cells: monolayer permeability and sensitivity to kinase inhibitors
| Non-5′HA-Verge-expressing cells | 5′HA-Verge-expressing cells | |||||||
|---|---|---|---|---|---|---|---|---|
| Treatment | Control | U0126 | SB | BIM | Control | U0126 | SB | BIM |
| Vehicle | 0.998 ± 0.03 | 0.998 ± 0.05 | 0.903 ± 0.05 | 0.973 ± 0.06 | 1.003 ± 0.02 | 0.996 ± 0.07 | 0.910 ± 0.07 | 0.980 ± 0.08 |
| PMA | 0.940 ± 0.08 | 0.927 ± 0.06 | 0.960 ± 0.08 | 0.880 ± 0.08 | 0.561 ± 0.12** | 0.631 ± 0.07** | 0.551 ± 0.10** | 0.860 ± 0.08 |
Normalized TER values are means ± SD in Ω. Each experiment represents the pooled TER values of two independent control (clones A and B) and two independent stable Verge-expressing (clones 38 and 41) cell lines (n = 3). RBE4 cells were grown to confluence on ECIS plates, and cells were rinsed with DMEM to remove serum and incubated in fresh DMEM for 30 min to achieve stable basal electrical resistance. Cells were then treated with vehicle, U0126 (10 μm), SB203580 (10 μm), or BIM (1 μm) for 30 min followed by PMA (100 nm) for 1 hr. Stimulation of Verge-expressing cells, but not control cells, with PMA leads to a significant decrease in TER. Pretreatment of Verge-expressing cells with BIM blocks the PMA-induced increase in permeability. Pretreatment with either U0126 or SB203580 produced no effect (**p < 0.05).