Barkley would miss the rest of the season if doctors confirm the injury.
The star back needed a cart to get from the sideline to the locker room to exit the field, and the team quickly ruled him out with a knee injury.
Barkley underwent X-rays and will have further evaluations Monday.
The 23-year-old had four carries for 28 yards before the injury.
Barkley had the second-worst rushing performance of his young career in Week 1, finishing the loss to the Pittsburgh Steelers with just six yards on 15 carries.
The 2018 No. 2 overall pick - who racked up 3,469 scrimmage yards and 23 touchdowns in his first two seasons - is the key cog of New York's offense.
If he's indeed lost for the year, it would be a devastating blow to the Giants' playoff hopes.