Guests include: Q-Tip, Usher, Carlos Santana, Chris Rock, Bruce Springsteen, Elvis Costello, Maya Rudolph, Solange, Shep Gordon, Steve Miller, "Weird Al" Yankovic, The Revolution, and Chaka Khan.

QLS also sometimes features public figures outside the music industry such as John Oliver, Bill Clinton, Stacey Abrams, and Michelle Obama.

[4][5] Questlove Supreme usually begins with a musical introduction, where the panelists participate in a "Shabooya Roll Call," an "age-old school bus chant" ad-lib format.

[6] Questlove hosts the show, with Team Supreme co-hosting, providing additional questions, reactions, jokes, and references.

QLS guests are asked about their early musical experiences, influences, and careers, with other off-script questions emerging based on the conversation.