The Williams sisters’ brief doubles comeback at the US Open ended in a loss in the opening round.
They lost to Lucie Hradecka and Linda Noskova of the Czech Republic while pairing up for the first time since 2018, likely for the last time.
The American greats have combined for 14 Grand Slam doubles victories.
After the competition, Serena Williams, 40, is scheduled to retire, while Venus, 42, is still unsure of her future plans.
Venus was eliminated in the first round of the women’s singles tournament, while Serena faces Australian Ajla Tomljanovic in the third round.
The sisters opened Thursday’s night session at Arthur Ashe Stadium by failing to convert two set points on their opponents’ serve at 5-4 after both pairs had earlier missed break chances.
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On their first set point in the ensuing tie-break, Hradecka and Noskova prevailed, and the 37-year-old Hradecka finished things off with a strong backhand down the line.
She continued to dominate the second set, securing an early break with Noskova, who is 20 years younger than she is. The Williams sisters held and ended a five-game losing streak shortly after holding a double break point for 4-0, but Noskova could not convert from close to the net.
They continued to break back, but on their second match point, Hradecka and Noskova broke their serve once more, so their efforts were in vain.
With victories at Wimbledon and the US Open in 2009, followed by victories at the Australian Open and French Open in 2010, the Williams sisters are still the most recent women’s doubles team to hold all four major doubles titles simultaneously.
While they also have three Olympic doubles gold medals, their most recent Grand Slam doubles victory came at Wimbledon in 2016, more than 17 years after their first at Roland Garros in 1999.