Hiking can cause discomfort and injuries at the lower body part. In order to prevent and reduce injuries, wearing knee supports is very important. However, studies related to knee supports are so limited in Taiwan that leads to our interests. This study aimed to explore the effects of wearing knee supports on different slopes on mountain hikers. A total of 12 male subjects participated in the experiment. Experimental design used the two-way randomized complete block experiment. Each subject was a block, and the variables included types of knee supports (knee sleeves, Velcro-type, strap, and no knee supports) and slopes (uphill, flat, and downhill). Dependent variables included heart rate (HR) and rating of perceived exertion (RPE). The results showed that (1) types of knee supports had significant effects on HR and overall RPE. HR and overall RPE of the knee sleeve group were significantly less than the group without knee supports, but had no significant differences between the Velcro-type and straps. (2) Slopes also had significant effects on HR and overall RPE. HR and overall RPE on uphill were significantly higher than on the flat and downhill, but there was no significant difference between the flat and downhill. (3) Types of knee supports and slopes had significant interaction on knee RPE and overall RPE. Overall, using knee sleeves on uphill and downhill, while using knee straps on flat, can reduce the burden and physical cost of the lower part.