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The group topped the bill on the Pyramid Stage on Friday (26 June) night, closing the first day of music at the event.
The band were promoted to the headline slot after original act Foo Fighters were forced to pull out due to frontman Dave Grohl breaking his leg.
Florence Welsh and the rest of her band performed a rousing set to thousands of fans at Worthy Farm, playing hits including ‘Dog Days Are Over’, ‘You Got The Love’ and ‘'Say Nothing'.
They also paid tribute to Dave Grohl, dedicating a cover of Foo Fighters' 'Times Like These' to him.
Kanye West is set to headline the Pyramid Stage on Saturday night, with The Who bringing the festival to a close on Sunday.
Watch a clip of Florence and The Machine’s set above, or watch the full thing on the BBC iPlayer.