Glucose glo assay kit

Growth defects associated with inhibited snf1 function were assessed by growing wild-type and mutant strains on YP media (2% yeast extract, 1% peptone, and 2% agar) with 2% sucrose and antimycin A to a final concentration of 1 μg/ml. Growth rates for yeast strains were determined on SC-Ura media with 2% glucose as a carbon source for 24 h incubation at 30° C with shaking. Growth rates of kcs1 deletion strains carrying wild-type KCS1 and the kcs1-S537A,S646A mutant were calculated as the slope (increase in culture optical density over time) from the previous time point. Mean growth rate was calculated from triplicate biological replicates, and the error bar indicates one standard deviation. Glucose levels in liquid cultures were measured using the Glucose-Glo assay kit (Promega). Glucose levels are shown as a mean percentage of starting glucose concentration with error bars indicating one standard deviation.

Glucose‐Glo™ Assay Kit (Promega) was used to determine glucose uptake according to the manufacturer's protocols. IL‐4‐primed M2 peritoneal macrophages from Elp3^Control and Elp3^ΔMye mice were plated at 2.5 × 10⁵ cells/ml into 24‐well plates and incubated for 24 h. Next day, cells incubation mediums and medium without cells (empty control) were collected and used for determination of glucose consumption. The luminescence was recorded by PerkinElmer Victor X3. The glucose consumption was calculated as the difference between the noncells medium value and cells incubation medium value.

Glucose uptake was measured with a Glucose-Glo™ assay kit (Promega, Madison, WI, USA). Lactate secretion was measured with Lactate-Glo™ assay (Promega). Measurements were normalized to cell number.

For measurement of lactate and glucose levels, lactate and Glucose-Glo™ assay kits were purchased from Promega (Madison, WI, USA). CCA cells were plated in 96 well plate in plain RPMI or DMEM media with no added growth factors overnight. They were then treated with plain EGM (containing 5% FBS), LPS-LEC-CM, plain RPMI/DMEM (containing no FBS), and CXCL5. After 24 h, the cells were treated with lactate and glucose detection reagents and lactate and glucose levels were measured in luminescent mode as per manufacturer’s protocol.

After 24 h of transfection, primary hepatocytes were washed with warm 1× PBS, and glucose-free DMEM (Gibco) supplemented with 100 mM sodium pyruvate was added in order to study cell-intrinsical glucose production. Stimulation with Fsk/Dex (10 µM/100 nM) with/without 100 nM insulin was performed for 24 h at 37 °C and 5% CO₂. Supernatant medium of the cells was collected to measure endogenous glucose production with Glucose-Glo Assay Kits (Promega) following the manufacturer’s instructions.