[2] KWU competes in 22 intercollegiate varsity sports, listed below:[3] The men's basketball team advanced to the Division II championship game six consecutive years (1998–2003), winning in 1999 and 2001.

Kentucky Wesleyan has an American football team, which competes in the Division II Great Midwest Athletic Conference.

[5] The Panthers primarily compete in the Great Midwest Athletic Conference (G-MAC), as a founding member since the 2013–14 academic year.

They also competed in the Kentucky Intercollegiate Athletic Conference (KIAC; now currently known as the River States Conference (RSC) since the 2016–17 school year) of the National Association of Intercollegiate Athletics (NAIA) from 1916–17 to 1954–55.

Kentucky Wesleyan's athletic teams have won eighteen (18) Great Lakes Valley Conference (GLVC) championships.