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The 14th-seeded Eagles (24-8) sizzled from 3-point range early and pushed Villanova to the brink of a monumental upset. They just didn't have enough left in their tired legs down the stretch, something the Wildcats exploited in their game-changing spurt.
The loss ended the Eagles' 13-game winning streak, the second-longest in the country.
Garrison Carr scored 22 points, Derrick Mercer had 17 and Brian Gilmore 16 for American, whose senior starters not only wanted to leave it all on the floor, they never wanted to leave the floor. Mercer played 40 minutes and Carr (37) and Gilmore (36) were right behind.
The Wildcats will play the UCLA-Virginia Commonwealth winner on Saturday.
For 30 minutes, American was poised to escape with the win in only the program's second NCAA tournament game. But Anderson and Cunningham, part of the winningest senior class in Villanova history, refused to let that happen.
Anderson hit a 3-pointer for a 58-55 lead with 6:21 left that brought the Wildcats all the way back from their big deficit. Cunningham followed with a layup and Villanova was shifting into another gear while the Eagles were tiring.
American built its lead on 3-pointers, going 8-for-15 in the first half, before fizzling the rest of the way.
They only went to the free-throw line five times, while Villanova made 26 of 29 attempts and chipped into the lead. Corey Fisher made four during Villanova's big run trim the gap to four, and Scottie Reynolds, a non-factor for most of the game, finally tied it for Villanova on a driving layup that made it 55-all.
The 3-point shot is the great equalizer in tournament play and American came prepared.
Carr nailed one in transition for a 24-22 lead and followed with another the next time down, the first of his four in the opening half.
Gilmore capped the half with a 3 from the top of the arc, sending American into the locker room with a 41-31 lead and the sound of the school band's rendition of "American Woman" following them into the locker room.
The scene had to look painfully familiar for coach Jay Wright and the Wildcats.
They were a No. 1 seed and in a similar situation three years in their first round game against No. 16 Monmouth. Monmouth hung around all game and trailed by seven with 6:22 left before the Wildcats pulled away in a 13-point victory at the Wachovia Center.
They did the same thing again not even 20 miles away from the suburban campus.
American wasn't all about the long ball, though. Gilmore put it on the court for a driving layup and the first basket of the second half. Mercer - think of New York Knicks dynamo Nate Robinson - split defenders in the lane for a bucket and a 45-31 lead.
The neutral fans, especially the ones wearing UCLA blue, joined the American supporters and made it sound like the entire Wachovia Center was pulling for the 17-point underdog.
So much for Villanova's home-court advantage.
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