The ad showed a 35-minute drill of burpees and rowing exercises.
At the bottom were the words: "And don't you dare lay down."
People on social media claim the sign is mocking the death of George Floyd at the hands of police last month.
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The shocking incident sparked Black Lives Matter protests across theworld.
One woman on Twitter said the sign was "unacceptable".
"The franchise needs to be pulled immediately as well as firing that trainer," she said.
"Absolutely disgusting."
Another said it was a "completely irresponsible, tone deaf workout".
Anytime Fitness was forced to apologise over the incident.
The co-founders and brand president of the company, which owns more than 4000 franchises across 50 countries, including Australia and New Zealand, said they were "shocked and devastated" over the sign.
In a separate update they said the trainer had been placed on leave and his employment status was under review.
They said the franchise owner and his staff volunteered to be the first to do anti-racism training the company announced last week.
