The Supernatural actors have teamed up once more for season 2 of Walker, with Ackles, 43, directing an episode for the latest season of The CW show, which Padalecki, 39, stars in and serves as executive producer for, Entertainment Weekly reported Thursday.
The project marks the first time the actors have collaborated since Supernatural wrapped after 15 seasons on The CW. The show aired from 2005 to 2020, and the two played brothers Dean and Sam Winchester.
In June, Padalecki claimed he only learned about Ackles' upcoming Supernatural prequel through social media, expressing shock following the announcement of the series.
"Dude. Happy for you. Wish I heard about this some way other than Twitter," Padalecki wrote to his friend. "I'm excited to watch, but bummed that Sam Winchester had no involvement whatsoever."
Shortly before Walker premiered in January, Padalecki told PEOPLE that he had expressed interest in reprising his Supernatural role if offered the opportunity.
"I certainly have it somewhere in the bottom of my heart and soul that I hope September 10 isn't the last time I ever played Sam Winchester," he said in reference to the final filming date.
Padalecki later cleared the air around his comment on Ackles' prequel post, noting that the two "had a great talk" and that "things are good" between them.
"We've travelled a lot of roads together, and sometimes those damn roads have bumps. Bumps don't stop us," the actor said after they spoke. "Once brothers, always brothers."
Ackles retweeted Padalecki, adding, "Love you @jarpad… Miss these talks. I forgot how much face time we always used to get. And I miss that too. I know you're busy…as am I, but you're still my brother. I miss you, pal."
"He's my brother — he has been for many years, and he always will be, no matter what," he told the newspaper. "He's spent more time with me on camera than anybody probably ever will, so he knows my strengths and weaknesses more than I do, and vice versa. I respect his opinion."
Season 2 of Walker premieres Oct. 28 at 8 p.m. EST on The CW.