The 'Somebody Told Me' hitmakers - who were playing the event for the fourth time - invited the comedian on stage at Hylands Park, Chelmsford, Essex, to introduce their 2006 hit single 'When You Were Young'.
He told the crowd: ''You have two options: You can call it a night, you might be fresher for work in the morning, or you could demand more?''
His first suggestion prompted boos from the audience but they were left cheering when he walked off stage with the returning band heading in the opposite direction following a short break at the end of their energetic greatest hits set.
That wasn't the Las Vegas band's only surprise of the night though as they also performed a superb cover of Iggy Azalea and Charli XCX's 'Fancy' resulting in drummer Ronnie Vannucci stepping out from behind his drum kit to pull off some suggestive dance moves while frontman Brandon Flowers sang.
But The Killers were not the only impressive act of the weekend as several other stars excelled at the festival.
Lily Allen got plenty of plaudits for her set on the main Virgin Media Stage but it was a wardrobe malfunction which stole the show as her plunging silver jumpsuit plunged a little too far leaving the bra-less star's assets on display for all to see.
Sam Smith continued his fantastic year by entertaining fans on the star-studded MTV Stage on Sunday, which also featured Rizzle Kicks, Tinie Tempah and Jason Derulo among other big names.
M People had got V Festival's Sunday off to the perfect start by rolling back the years with a greatest hits set and Train, Chic and Rudimental followed suit later in the day.
On Saturday (17.08.14), Ed Sheeran wowed the crowd with a selection of hits from his two albums and he later received a somewhat unexpected shoutout from Saturday night's headline act Justin Timberlake.
Following his performance of 'Cry Me A River', Justin told the crowd: ''Brother from another red-headed mother Ed Sheeran - he's a bad motherf**ker!''
Other Saturday performers at the festival included Example, Kaiser Chiefs and Bastille.
A number of stars also made it into Virgin Media's Louder Lounge area including 'Skins' beauty Kaya Scodelario, Katherine Jenkins and The Saturdays' Vanessa White.
And when people weren't living it up in the Louder Lounge or taking in the fine selection of acts performing at the music extravaganza, some were having a fantastic time at the Lucozade Energy YES Arena.
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