D glucose saccharin solution

During the 11^th week of the chronic exposure, mice were weighed and briefly exposed to a D-glucose:saccharin solution (w/v; 3.0%/0.125%; Sigma-Aldrich; St. Louis, MO) immediately prior to 6h exposure. After CR and MCS exposures, mice were placed singly per metabolic cage (Harvard Apparatus; Cambridge, MA) with glucose:saccharin solution drinking water but without food for urine collection (in graduated cylinders surrounded by 4°C water-jacketed organ baths). Urine was collected in 1h increments up to 3h post-exposure followed by an overnight urine collection during which mice were provided glucose/saccharin solution as well as food (Conklin et al., 2017b (link)). Collected urine samples were centrifuged (1,800xg, 5 min; to pellet any feces or food particles) before being decanted and stored at −80°C.

On day 1, mice were weighed and briefly exposed to D-glucose/saccharin solution (w/v; 3.0%/0.125%; Sigma) immediately prior to HEPA- and charcoal-filtered air exposure (6h), and then placed singly per metabolic cage (Harvard Apparatus) with glucose/saccharin solution in drinking water without food to collect urine (in graduated cylinders in 4°C water-jacketed organ baths) for a continuous 3h collection post-exposure. After urine collection, mice were placed in home cages overnight with food and water per normal housing arrangements. On day 2, the same mice were exposed to aerosols of KY Reference cigarette mainstream smoke (3R4F cigarettes: 12 cigarettes/6 h) or an e-cig (blu®; Classic Tobacco, CT; Magnificent Menthol, MM; 13–16 mg nicotine; 4h) and then placed singly per metabolic cage with glucose/saccharin solution to collect urine (without food) in 1 h increments over the 3 h post-exposure followed by an overnight (O/N; with food) urine collection. Urine samples were collected, decanted, and stored at −80 °C (Conklin et al. 2017 (link)).