| 04 Jun 25 |
r09        | Charlotte Independence IICharlotte Ind | 0 - 1 | Salem CitySalem City |
| 04 Jun 25 |
r09        | Wake FCWake FC | 2 - 3 | North Carolina FC U23North Carolin |
| 04 Jun 25 |
r09        | Lehigh Valley UnitedLehigh Valley | 1 - 0 | Real Central New JerseyReal Central |
| 04 Jun 25 |
r09        | FC Motown IIFC Motown II | 1 - 0 | New Jersey CopaNew Jersey Co |
| 04 Jun 25 |
r09        | Asheville City SCAsheville Cit | 1 - 0 | Southern Soccer AcademySouthern Socc |
| 04 Jun 25 |
r09        | Columbus UnitedColumbus Unit | 2 - 0 | Apotheos FCApotheos FC |
| 04 Jun 25 |
r09        | Athens UnitedAthens United | - | Birmingham Legion 2Birmingham Le |
| 05 Jun 25 |
r09        | Fort Lauderdale UnitedFort Lauderda | 1 - 2 | Sarasota ParadiseSarasota Para |
| 05 Jun 25 |
r09        | Westchester FlamesWestchester F | 1 - 3 | Manhattan SCManhattan SC |
| 05 Jun 25 |
r09        | AHFC RoyalsAHFC Royals | 1 - 0 | AC Houston SurAC Houston Su |
| 05 Jun 25 |
r09        | Boston CityBoston City | 0 - 1 | AC ConnecticutAC Connecticu |
| 05 Jun 25 |
r09        | GFI AcademyGFI Academy | 0 - 0 | Corpus Christi FCCorpus Christ |
| 05 Jun 25 |
r09        | Twin City ToucansTwin City Tou | 2 - 4 | Lonestar SCLonestar SC |
| 05 Jun 25 |
r09        | Tennessee SCTennessee SC | 2 - 0 | East Atlanta Dutch LionsEast Atlanta |
| 06 Jun 25 |
r09        | Reading UnitedReading Unite | 0 - 0 | Ocean City Nor'eastersOcean City No |
| 07 Jun 25 |
r09        | Western Mass PioneersWestern Mass | 4 - 1 | New England FCNew England F |
| 07 Jun 25 |
r09        | Monterey Bay FC 2Monterey Bay | 1 - 1 | San Francisco GlensSan Francisco |
| 07 Jun 25 |
r09        | Christos FCChristos FC | 2 - 3 | Virginia Beach UnitedVirginia Beac |
| 08 Jun 25 |
r09        | Des Moines MenaceDes Moines Me | 10 - 1 | Springfield FCSpringfield F |
| 08 Jun 25 |
r09        | Albion SC ColoradoAlbion SC Col | 1 - 1 | Utah UnitedUtah United |
| 08 Jun 25 |
r09        | Colorado StormColorado Stor | 0 - 1 | Flatirons RushFlatirons Rus |
| 08 Jun 25 |
r09        | Academica SCAcademica SC | 2 - 2 | San Francisco City FCSan Francisco |
| 08 Jun 25 |
r09        | Lane UnitedLane United | 3 - 1 | Midlakes UnitedMidlakes Unit |
| 08 Jun 25 |
r09        | Snohomish UnitedSnohomish Uni | 2 - 1 | FC OlympiaFC Olympia |
| 08 Jun 25 |
r09        | Bigfoot FCBigfoot FC | 2 - 1 | Portland BangersPortland Bang |
| 08 Jun 25 |
r09        | RKC Third CoastRKC Third Coa | 4 - 1 | Chicago City Dutch LionsChicago City |
| 08 Jun 25 |
r09        | Union FC MacombUnion FC Maco | 1 - 2 | Oakland CountyOakland Count |
| 09 Jun 25 |
r09        | Virginia MaraudersVirginia Mara | 0 - 4 | Northern Virginia FCNorthern Virg |
| 10 Jun 25 |
r09        | Northern IndianaNorthern Indi | 0 - 0 | AFC Ann ArborAFC Ann Arbor |
| 10 Jun 25 |
r09        | West Virginia UnitedWest Virginia | 0 - 1 | Lexington SC IILexington SC |
| 10 Jun 25 |
r09        | Cleveland ForceCleveland For | 0 - 1 | Toledo VillaToledo Villa |
| 10 Jun 25 |
r09        | Lansing City FootballLansing City | 0 - 5 | Flint City BucksFlint City Bu |
| 11 Jun 25 |
r09        | Chicago City Dutch LionsChicago City | 1 - 2 | Minneapolis CityMinneapolis C |