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3uM of the 5-Aza deoxy derivative, 5-dAza-C, were treated with BRL-3 A for 36hr, and then harvested for indicated experiments.
For indicated experiments (those involving LPA treatment), cells were serum-starved by incubation in DMEM in the absence of FBS for 18 24 h after monolayers reached confluence.
Mice were bred and housed in our mouse facility (Chronobiotron, UMS 3415, CNRS, Strasbourg) and tested during the light phase (ZT2 ZT10) of their light/dark cycle except for indicated experiments.
After centrifugation and resuspension in α-MEM, the cell number was determined with a Cedex automated cell counter (Roche Diagnostics GmbH, Mannheim, Germany), and macrophages were seeded at a density of 1.5 × 10 cells/cm for indicated experiments.
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The bar chart in Fig. 7A shows that our model indeed produces nanoswitches, which are fully active even for small stimuli, as indicated by experiments [27].
For experiments indicated in Fig. 8, 2.5 µ m d-tubocurarine chloride was used together with 1 µg/mL tetraethylrhodamine-α-bungarotoxin The procedures described above were approved by the Institutional Animal Use and Care Committee at Grinnell College.
As a result, this indicates for experiments 1 5 that the starting temperature for DO/HDO via CO release is ≥230 °C.
The emulsions were kept in syringes, placed under light (30 35 μE/m/s), and incubated for lactate production as indicated for each experiment.
HeLa data were recorded using 75% normalized collision energy (CE) for HCD, whereas the HCD collision energy was varied as indicated for the experiments using the protein mixture.
Differences in timeline are indicated for specific experiments below.
Cells were treated, as indicated for individual experiments.
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