Abstract
We present a protocol to quantify the response of both normal and mutant Arabidopsis seedlings to gravity and simulated microgravity under earth-normal gravity conditions. We describe the steps to simulate microgravity using a three-dimensional (3D) clinostat, which changes the rate and direction at random and consistently rotates the axis horizontally and vertically to counteract the standard gravity at the Earth's surface. We then detail the gravity stimulation experiment, followed by the assessment of root responses using ImageJ-based analysis. For complete details on the use and execution of this protocol, please refer to Xu et al. (2022).(1).