The Upper Room headquarters in Nashville, Tennessee is centered around the Upper Room Chapel, which hosts thousands of visitors each year. Speakers from different denominations and countries take part in services (held regularly on Wednesdays), and visiting choirs from Nashville and elsewhere provide special services of music.

The focal point of the Upper Room Chapel is a woodcarving of Leonardo da Vinci's painting. The Last Supper, sculpted by Ernest Pellegrini. The woodcarving is an extraordinary work of art, created by more than fifty people over fourteen months' time. The carving is seventeen feet wide and eight feet high and, when viewed from the rear of the chapel, seems to have a depth of many feet, almost as if there were a room within the carving (although the greatest depth in the carving is only eight inches). The chancel of the chapel mirrors the carving, with the ceiling, tapestries, and altar table designed to reflect the setting in Pellegrini's work.

The pulpit in the chapel, with its winding stair and canopy, has features of several significant pulpits: City Road Church in London, made famous by John Wesley; St. George's Church in Philadelphia; and St. Philip's Church in Charleston, South Carolina, where Wesley preached when he was a missionary in Georgia. The front of the pulpit bears the Chi Rho, said to be the oldest monogram of Christ.

Visitors also enjoy the Upper Room Museum, whose permanent collection reflects the international, interracial, and interdenominational nature of The Upper Room. In December and January, the museum exhibits its collection of nativity scenes, with over one hundred renditions from cultures around the world. From Ash Wednesday through April, guests can view a special collection of more than seventy Ukrainian eggs. Hand-designed by Cheryl Christensen, the eggs tell the story of Easter and celebrate the beauty of spring using Christian symbols.