Overall, Twitter's stream of The New York Jets facing The Buffalo Bills on Thursday reached 2.1 million viewers, with an average of 243,000 watching the game at any given moment, according to the US National Football League.
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About 200,000 more people tuned in for a pre-game show.
People watched an average of 22 minutes of game action, the NFL said.
The event was the first of 10 Thursday night NFL football games set to stream for free on the San Francisco-based one-to-many messaging platform this season.
The Jets defeated the Bills by a score of 37 to 31.
The alliance was also part of an NFL effort to adapt to a viewership shifting away from traditional television or cable outlets to online platforms.
The league late last year teamed with Yahoo.com for the first global stream of an NFL game, attracting millions of viewers during the test.
Twitter accounted for the bulk of the "digital" audience for the game Thursday, according to figures released by the NFL.
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The game was also broadcast on CBS Television Network, with 15.7 million people watching across all platforms, according to the NFL.
Twitter reportedly paid $10 million dollars for rights to stream Thursday night games, and stands to make money from video ads woven into broadcasts.
Twitter this week announced a new application that will offer free content on major streaming TV services.
The new app for Apple TV, Amazon Fire TV and Microsoft's Xbox One will allow anyone with these devices to get content from Twitter's media partners.
That will include the NFL games, Bloomberg News and others.
"These devices will bring Twitter's live streaming video experience to life on the TV screen," Twitter chief financial officer Anthony Noto said when the apps were introduced.
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Earlier this week Twitter said it would be offering users live-streamed video programs on technology, media and business as the social network seeks to broaden its audience.
The new service will be in partnership with the live and on-demand video news network Cheddar.
The moves come as Twitter is struggling to keep pace with faster-growing social networks and expand beyond its core base of users to move to profitability.
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