The sweet moment took place when Lopez's 11-year-old daughter, Emme, joined the women on stage.
The ladies took the stage for the Pepsi Super Bowl LIV Halftime Show on Sunday night, where they performed a medley of their hits including "Whenever, Wherever," "Hips Don't Lie," "Jenny From the Block" and "Get Right."
It was when Lopez's 11-year-old daughter, Emme, joined her mom on stage to sing "Let's Get Loud," though, that the women paid tribute to the late NBA star, who died earlier this month in a helicopter crash along with eight others, including his 13-year-old daughter, Gianna.
In an overhead shot of the performance, fans spotted the tribute, which featured a cross that lit up yellow and purple, the colors of the Los Angeles Lakers where Bryant spent his 20-year career.
In a press conference prior to the big game, Shakira and Lopez teased the planned tribute.
"We'll all be remembering Kobe on Sunday and we'll be celebrating life and celebrating diversity in this country," Shakira said. "I'm sure he would be very proud to see the message that we're going to convey on stage that day."
Bryant was also remembered prior to kick off when the San Francisco 49ers and Kansas City Chiefs lined up at the 24-yard line -- a nod to Bryant's number -- for a moment of silence. Chants of "Kobe!" broke out following the quiet moment.
Players also honored the late athlete individually, with the Chiefs' Demarcus Robinson wearing one purple and one yellow shoe, Dante Pettis of the 49ers featuring the phrase "Mamba Out" on his cleats, and 49ers receiver Emmanuel Sanders' game-day shoes reading "Rest in Peace" with number "24" at the collar.
Watch the video below for more on the halftime show.