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Jimmy Kimmel Live! was pre-recorded at El Capitan Theatre in Hollywood and has now begun airing on ABC in the United States at 11.35pm PST (1.35pm AEST) today.
The host was treated to a standing ovation as he arrived on stage, before he launched into an emotional speech about Kirk and his suspension.
The host began his show with a montage of news clips covering his sensational suspension.
Kimmel was then seen wearing a bear suit while comedian Guillermo Rodriguez dressed as a banana.
As he kicked off his emotional monologue, Kimmel’s voice broke as he directly addressed the comments he made about Kirk.
”It was never my intention to make light of the murder of a young man,” Kimmel said.
“I don’t think there’s anything funny about it.”
Kimmel said it was not his intention to “blame any specific group for the actions” of Kirk’s alleged killer.
“It was obviously a deeply disturbed individual,” he said.
“That was really in the opposite of the point I was trying to make, but I understand that to some that felt either ill-timed or unclear, or maybe both.
“And for those who think I did point a finger, I get why you’re upset.
“If the situation was reversed, there’s a good chance I’d have felt the same way.”
Kimmel then explained he doesn’t believe the man who shot Kirk represents anyone.
He described him as a “sick individual who believed violence was a solution”.
”And also selfishly, I am a person who gets a lot of threats. I get many ugly and scary threats against my life, my wife, my kids, my co-workers because of what I choose to say,” Kimmel continued.
“And I know those threats don’t come from the kind of people on the right who I know and love.”
He praised Kirk’s widow Erica for publicly forgiving her husband’s alleged killer.
“Erica Kirk forgave the man who shot her husband. She forgave him. That is an example we should follow,” he said.
“A selfless act of grace, forgiveness from a grieving widow. It touched me deeply and I hope it touches many.”
As his attention turned to Donald Trump, Kimmel said the president’s attempt at censoring him had backfired.
“He tried his best to cancel me. Instead, he forced millions of people to watch the show,” he said.
“He might have to release the Epstein files to distract us from this now.”
Kimmel also poked fun at Trump’s latest headlines, a running theme in his monologues.
This time, it was about the president’s announcement linking Tylenol use during pregnancy to autism in children.
“Follow the advice of Donald Trump and you too can look like a glazed ham with deep-vein thrombosis,” he joked.
Kimmel’s monologue marked his first public comments since being pulled off-air last week.
The show was sidelined after comments the host made about Kirk’s death.
He said in his September 15 monologue that the “MAGA gang” were ”desperately trying to characterise this kid who murdered Charlie Kirk as anything other than one of them”.