For GTT and PTT, 12- to 16-week-old male GF and SPF mice were fasted overnight for 14 hours, starting at ZT12. For ITT, mice were fasted for 5 hours, starting at ZT22. Baseline blood glucose was measured via tail snip using a hand-held glucometer. Mice were administered an oral bolus of 20% dextrose (Hospira) in sterile water solution (2 g/kg body weight), an intraperitoneal injection of sterile-filtered sodium pyruvate (Sigma-Aldrich) in PBS (2 g/kg body weight), or an intraperitoneal injection of 0.1 U/mL Humulin R Insulin (Eli Lilly) in PBS (1 U/kg body weight). For GTT and PTT, blood glucose was measured at 1, 30, 60, and 120 minutes after gavage or injection. For ITT, blood glucose was measured at 15, 30, 60, 90, and 120 minutes after injection. During GTT, insulin levels were determined at baseline, 30, 60, and 120 minutes in blood collected in heparin-coated microvette tubes (Sarstedt) using the Ultra-sensitive Insulin ELISA (ALPCO). AUC was calculated using GraphPad Prism v9.
Heparin coated microvette tubes
Manufactured by Sarstedt
Sourced in Germany
Heparin-coated microvette tubes are a type of blood collection tube designed for immediate plasma separation. The tubes are coated with heparin, an anticoagulant, to prevent blood clotting during sample collection and processing.
Automatically generated - may contain errors
2 protocols using heparin coated microvette tubes
1
Glucose and Insulin Tolerance Tests
2
Creatine Kinase Measurement Protocol
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For all cohorts, creatine kinase (CK) levels were measured at baseline, three and five weeks after treatment initiation, and prior to sacrifice. Hereto, blood was taken via a small cut in the tail vein and collected in heparin-coated microvette tubes (Sarstedt, Numbrecht, Germany). Samples were kept on ice and plasma was obtained after centrifugation at 13,000 rpm for 5 min at 4 °C. Plasma CK levels were measured on the Reflotron Sprint system with CK Reflotron strips (Roche, Diagnostics, Basel, Switzerland).
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