Jodie Foster, Michael J. Fox and Keegan-Michael Key were among the stars who turned out for the rally at United Talent Agency's headquarters in Beverly Hills.
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"We'll never bow down to division, to cruelty, to fear," Fox said.
The "United Voices" rally was aiming to express the creative community's support for freedom of speech and artistic expression, while standing against policies of exclusion.
In a taped appearance, special guest Asghar Farhadi spoke to the crowd from Iran.
The director won an Oscar for best foreign language film in 2012 and his new movie "The Salesman" is nominated for best foreign film at this weekend's Academy Awards. Farhadi decided to boycott the ceremony after the announcement of President Trump's travel ban, which prohibited travelers from seven Muslim-majority countries.
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"I hope this unity will continue and spread to fight other injustices," Farhadi said.
Among other participants at the event were California Lt. Gov. Gavin Newsom, singer Ben Harper, model Emily Ratajkowski and actors Megan Mullally, Nick Offerman and Kristen Wiig.
The event was also a fundraiser, with UTA pledging to donate $250,000 to the ACLU and the International Rescue Committee. An additional $70,000 was raised online.