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The US government agency that regulates television says it is opening an investigation into Disney's diversity and inclusion practices, in the latest sign of pressure being applied to media firms.
Federal Communications Commission (FCC) chairman Brendan Carr sent a letter to Disney on Friday notifying the firm and its ABC News unit of the plan.
He said the move had been prompted by concerns that the company was promoting diversity "in a manner that does not comply" with government regulation.
A spokesperson for Disney said the company is reviewing the letter.
"We look forward to engaging with the commission to answer its questions," a Disney spokesperson said in a statement.
The FCC's investigation into Disney also comes amid a wider crackdown on DEI practices by the Trump administration, with impacts felt beyond the US.
French companies with US government contracts received a letter from the American embassy in France this week, asking them to sign on and comply with Trump's executive order banning DEI programmes.
The order, the letter said, "applies to all suppliers and service providers of the US government, regardless of their nationality and the country in which they operate."
In a letter addressed to Robert Iger, the chief executive of Disney, FCC chairman Carr said he wants to ensure that the media company "ends any and all discriminatory initiatives in substance, not just name".
He added: "I want to determine whether Disney's actions - whether ongoing or recently ended - complied at all times with applicable FCC regulations."
Carr has been a member of the FCC since 2017 and was named to lead the agency by Trump in November.
Since being appointed to the post, he has ramped up scrutiny of media firms, launching probes of NPR and PBS and demanding information from Big Tech companies, including Apple and Google, about their use of services that influence how news articles get ranked.
The FCC also announced investigations of Verizon and Comcast and its media unit, NBCUniversal over their diversity, equity and inclusion (DEI) initiatives.
Disney, best known for its cartoon classics and theme parks, made changes to its DEI policies earlier this year.
"While I have seen reports that Disney recently walked back some of its DEI programs, significant concerns remain," Carr wrote in the letter.
"I want to ensure that Disney and ABC have not been violating FCC equal employment opportunity regulations by promoting invidious forms of DEI discrimination," he said.
Carr said he wanted information about rules governing diversity representation in its characters, among other initiatives.
The inquiry is not the first time Disney has been a political target.
Last year, it agreed to pay $15m to settle a defamation lawsuit brought by Donald Trump after a star ABC anchor falsely said the president had been found "liable for rape".
A New York jury had previously determined Trump was liable for "sexual abuse", which has a specific definition under New York law.
Republican Florida Governor Ron DeSantis also spent years feuding with Disney, after the company criticised a state law, known as "Don't Say Gay", that restricted the teaching of sexuality in schools.
Disney has previously been a target of conservatives for allegedly embracing "woke" messages in its films.
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