Venus beat her younger sister last night in the third round of the BNP Paribas Open in a straight-sets 6-3 6-4 win.
Serena was making her competitive comeback after giving birth to her baby daughter, Alexis Olympia, in September.
Venus closed out the 29th career meeting between the two on her second match point as Serena sailed a forehand long to end the third round match.
Venus prevails in too-early sisters clash
But the former world number one was upstaged by a red-hot Venus in straight sets as the seven-time major victor reached the fourth round by beating Serena for the first time since 2014, despite a late wobble which saw her save a break point before moving through.
Venus's victory also comes 17 years after an ugly booing incident led to a 14-year boycott of the tournament by the sisters.
Serena, unseeded in the wake of her long layoff, admitted she would have preferred not to come up against her sister so soon in the draw. You know, she just did everything great.
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But when the siblings face off under the lights in Indian Wells, California, it will still be a hugely significant occasion for the Americans who have combined to win 30 majors and transcended tennis.
"I always know that it's never over until it's over", she said.
Serena made a decision to return in 2015, whilst Venus came back to Indian Wells in 2016.
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Talking about competing in the upcoming Grand Slams tournament, Serena was quoted as saying, "If I want to play in those grand slams and play well then now is the flawless time to start". "So it is what it is", said Serena.
"That's totally gone out of my mind", she said. Venus enjoyed a resurgent 2017 season, but she began this year with losses to Angelique Kerber in Syndey and Belinda Bencic at the Australian Open. In fact, it's the first time they've met before the Round of 16 since their very first head-to-head matchup more than 20 years ago.
"She gave me the trophy because I was so upset".
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The older Williams took all the momentum into the second set, breaking Serena in both of her opening service games for a 3-0 lead before Serena struck back, reducing the gap to just a single break in game four after pouncing on a weak Venus second serve. "I don't think my dad wants me to ever stop playing", the 37-year-old, seven-time Grand Slam champion said.