Electrophysiological recordings were performed from thalamic slices maintained in warmed ECS (30 °C), using an Axopatch 200B amplifier (Molecular Devices, Sunnyvale, CA, USA).
Animals were allowed to recover from anesthesia in a cage maintained in a warm room.
No difference in body mass was detected between animals maintained in the warm (figure 2 b; F = 1.755, p = 0.182).
After recovery from anesthesia, mice were maintained in a warm recovery cage for 1 h and returned to home cages.
The mice were allowed to recover from anaesthesia in a cage maintained in a warm room and were administered analgeics (buprenorphine, 10 μg/kg) post surgery.
Anesthesia was induced with a slow intravenous 10 mg/kg [4.54 mg/lb] injection of propofol (Fresenius Kabi, Isola della Scala, Italy) [ 50] with the animals maintained in a warm room (28°C [82.4°F]) for 24 hours before the procedure.
The anesthetized animals were maintained in side position on the warming pad throughout the MRI session.
After the first incubation, monocytes were washed with warm PBS and maintained in RPMI supplemented with 10% pooled human blood serum for 5 days (medium was refreshed at day 3).
Briefly, slices (400 µm) were maintained in oxygenated (95% O2 / 5% CO2), warmed (30°C) artificial cerebrospinal fluid (ACSF) containing 124 mM NaCl, 4.4 M KCl, 25 mM NaHCO3, 1.0 mM NaH2PO4, 2.0 mM CaCl2, 1.2 mM MgSO4, and 10 mM glucose, and allowed to recover for at least 2 hr prior to the start of an experiment.
FLC expression is repressed by vernalisation (extended cold) and this repression is maintained in a PRC2-dependent manner following return to warm growing conditions [ 11, 30].
