8, 20 Wild-type hepatocytes were rendered necrotic by incubation at 60°C for 60 minutes as described.
Microarray slides were pre-hybridized in 20% formamide, 5× Denhardt's, 6× SSC, 0.1% SDS, and 25 μg/ml tRNA for 45 minutes as described by Band et al. [ 19].
The membranes were next incubated in a solution of 2,4-dinitro-phenylhydrazine (0.5mM) in 2N HCl for exactly 5 minutes each, as described by Robinson et al. (6).
Baroreflex sensitivity expressed as bradycardic (BR) and tachycardic (TR) responses in beats per minute per millimeter of mercury, as described elsewhere.
Baroreflex sensitivity was expressed as bradycardic response (BR) and tachycardic response (TR) in beats per minute per millimeter of mercury, as described elsewhere [ 10].
Organotypic cultures at 10 DIV were deprived of oxygen and glucose for 30 minutes as described above.
For activation of the HOG pathway, yeast cells were treated with 0.4 mM NaCl for 30 minutes as described [14].
All samples were treated with RNAse-free DNase (QIAGEN, Valencia, CA, #79254) for 20 minutes as described in the manufacturer's protocol.
For experiments with inactivated RSV, the virus was exposed to an UV light source for 20 minutes as described previously [43].
Live embryos were dechorionated with forceps and then incubated in E3 buffer containing 5 µg/ml acridine orange (Sigma) at room temperature for 10 minutes as described [27].
Culture samples were removed at t = −1, 10, 20, 30, 40, 50, 60 and 80 minutes as described above and total RNA isolated using RNeasy Total RNA Isolation Kit (Qiagen).
