Cycloheximide treatment did not prevent aldosterone-induced increases in mRNA levels during the first hour (Supplementary material online, Figure S 2 ), suggesting that aldosterone activated pre-existing regulatory factors.
This increase required the presence of the drug for 2-4 hours, was sustained for at least 30 hours (Supplementary Figure 1B), and depended on both cytoplasmic and mitochondrial translation (i.e. was inhibited by addition of either cycloheximide or chloramphenicol; data not shown).
These data showed that by 24 hours there was no significant improvement of parameters as measured at 6 hours as a result of CM_128 administration begun at 6 hours (Supplementary Figures 3 and 4), confirming delay in therapy to be disadvantageous; although CM_128 appeared to prevent these parameters from increasing.
The acute administration of CL316243, BRL37344 and ICI215,001 at 0.1, 0.5 and 1 mg/kg body weight induced significant dose-response reductions of cumulative food intake over 24 hours in rats that had been previously food-deprived for 24 hours (supplementary material Fig. S1A C).
She is on supplementary oxygen 24 hours a day and spends most of her time in hospital having strong antibiotics delivered into her bloodstream, to help keep her alive while she waits for her transplant.
TUNEL-positive cells were noted to extend to the midbrain of embryos treated with nestin MO for 49 hours (Supplementary Fig. S2).
To determine the time dependence of the drug effect of mitomycin C, cells were treated with desferrioxamine, which caused a time-dependent increase in HIF-1α protein expression with a maximal effect at approximately 10 hours (Supplementary Fig. S2c).
Instead, GFP-expressing Photorhabdus appeared to be cleared from the gut within 24 hours (supplementary material Fig. S7).
Sma expression increased during development, with a peak level at about 12 hours (supplementary material Fig. S7).
TiPM were stimulated with muramyl dipeptide (MDP), Pam3 and HkEc for 24 hours (supplementary material Fig. S5E).
