At least one club from the second-tier USL will advance to U.S. Open Cup quarterfinals as Louisville City were drawn against Nashville SC in the round of 16.
U.S. Soccer conducted the draw on Friday, the morning after Nashville, whose owners were granted an MLS expansion team last year, advanced by defeating the Colorado Rapids.
Nashville SC will travel to face last season's USL champions Louisville City, which ousted the New England Revolution on Tuesday night.
The draw was split into four geographic regions, so the winner will advance to play either Atlanta United or the Chicago Fire, who will meet in Georgia later this month.
In the West Region, the Sacramento Republic, the only other non-MLS club to reach the round of 16, will play at LAFC, while the Portland Timbers host the LA Galaxy.
The Houston Dynamo will be at home against Minnesota United in the Midwest Region while FC Dallas travels to Sporting Kansas City.
And in the East, the Philadelphia Union will host the New York Red Bulls while Orlando City visits D.C. United.
Atlanta, LAFC, Houston and Philadelphia will host quarterfinal games in July should they advance.