Project protocol - Contents

• Workflow and sampling
• Equipment and supplies
• Reagents and solutions
• Procedure: Voluntary exercise
• Data

Workflow and sampling

Step	Procedure	Equipment	Data collected
1	Voluntary wheel running	Standard laboratory cages with attached running wheels	Distance traveled, duration, average speed
2	Measure food intake and assess body composition	Scales and MRI	See McMullan3

Equipment and supplies

• Running wheels (1.1 m circumference) (Lafayette Industries, Lafayette IN)
• Automated Activity Wheel Monitoring Program (AWM, Lafayette Industries, Lafayette IN)

Procedure: Voluntary exercise

1. Mice are individually housed in standard laboratory cages with attached running wheels.
2. Mice are allowed ad libitum access to running wheels for a month (28 days).
3. Voluntary wheel running is recorded continuously in 1-min intervals (as number of revolutions) using software from the manufacturer.

Data collected by investigator

• Daily distance traveled (total revolutions x 1.1 m)
• Daily duration (cumulative 1-min intervals in which at least 1 revolution is recorded)
• Daily average speed (total revolutions/total duration)