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Abstract Background Anti-gravity treadmills are used to decrease musculoskeletal loading during treadmill running often in return to play rehabilitation programs. The effect different gradients (uphill/downhill running) have on kinetics and spatiotemporal parameters when using an AlterG® treadmill is unclear with previous research focused on level running only. Methods Ten well-trained healthy male running athletes ran on the AlterG® treadmill at varying combinations of bodyweight support (60, 80, and 100% BW), speed (12 km/hr., 15 km/hr., 18 km/hr., 21 km/hr., and 24 km/hr), and gradients (− 15% decline, − 10, − 5, 0, + 5, + 10 + 15% incline), representing a total of 78 conditions performed in random order. Maximum plantar force and contact time were recorded using a wireless in-shoe force sensor insole system. Results Regression analysis showed a linear relationship for maximum plantar force with bodyweight support and running speeds for level running (p |