The nozzle temperature before deposition was maintained at 32 °C in order to extrude the melted state of the sample as the flow behaviour curves indicated that the melting of chocolate started between 28 °C to 30 °C.
Cells were pre-warmed to 37°C before analysis and were maintained at 37°C during event collection on a Becton Dickinson (BD) LSR II Flow Cytometer (Beckton Dickinson).
The laser power at the sample was measured before each run and was maintained at < 0.4 mW.
The soil water contents before and after watering were maintained at 54 60, 45 50, 34 40 and 22 25 % for the treatments, respectively.
For live-cell imaging, transfected cells were replated in Lab-Tek chambered coverglasses before imaging, and cells were maintained at 37 °C with 5% CO2.
Transfected cells were re-plated in Lab Tek Chambered coverglass (NUNC) the night before imaging, and cells were maintained at 37 °C with 5% CO2 in a LCI chamber (LCI, Seoul, Korea).
For each treatment, incubation temperature before and after flooding was maintained at 23, 28 and 33°C (i.e., 8°C higher than the respective flooding temperature) to approximate changes in soil temperature associated with heavy rainfall events recorded at a depth of 10 cm in the field (APLC, unpublished data).
The chambers were then sealed and placed in incubators at temperatures of 20, 25 or 30°C for 29 45 h (with shorter duration at higher temperatures, to compensate for higher rate of O2 consumption), such that the ΔPO2 in the chamber before and after the experiment was maintained at <1% (actually from 0.2 to 0.7) to avoid the potential stress of oxygen suppression on the embryos.
The temperature of all stored samples was maintained at 0 4 °C until immediately before analysis.
