TABLE 1.
Relative membrane expression of AQP4-GFP in response to hypotonic stimulus in the presence of inhibitors
Translocation of AQP4-GFP in HEK293 cells in the presence of various inhibitors, measured by confocal microscopy and subsequent image analysis, is shown. Cells were imaged before and after reduction of the extracellular osmolality to 85 mosm/kg of H2O. Each RME value is an average over three independent experiments, with each experiment analyzing at least 3 cells/micrograph and 3 line profiles/cell. p < 0.05 was considered statistically significant (*). Abbreviations: PKCi, PKC inhibitor; PKAi, PKA inhibitor.
| Inhibitor/control media | Isotonic RME (± S.E.) | Hypotonic RME (± S.E.) | Mean % of area change (± S.E.) | Translocation |
|---|---|---|---|---|
| Untreated control | 28.77 (6.78) | 78.84 (1.84)* | 145.19 (1.11)* | Yes |
| Non-specific kinase inhibitor (hypericin) | 21.85 (6.80) | 38.22 (5.58) | 103.50 (2.05) | No |
| PKC (Myr-PKCi) | 27.27 (4.73) | 55.28 (3.97)* | 141.00 (2.05)* | Yes |
| PKA (Myr-PKAi) | 25.62 (5.16) | 33.66 (6.62) | 102.97 (5.21) | No |
| Calcium-free media | 26.69 (6.03) | 32.16 (4.48) | 111.60 (1.92) | No |
| Calmodulin inhibitor (trifluoperazine) | 27.23 (7.11) | 31.40 (5.91) | 108.61 (5.47) | No |
| Calmodulin inhibitor (W7) | 16.86 (8.43) | 22.11 (8.14) | 110.31 (14.21) | No |