Venus Williams' valiant efforts at the Rothesay Classic in Birmingham ended in a second-round defeat by Jelena Ostapenko.
The 43-year-old pulled off her best victory for nearly four years with a three-hour win over Camila Giorgi in the first round on Monday despite struggling with a right knee problem.
She had the joint strapped up again on Thursday and took a long medical timeout during the second set for treatment to her right thigh in what looked like being a straight-sets loss.
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But Williams saved a match point at 3-5 in the second set and then reeled off three straight games as second seed Ostapenko lost her rhythm.
When she moved 2-0 ahead in the deciding set with evening drawing in, it appeared Williams might be on course for back-to-back wins for the first time since 2019. But Ostapenko, a former Wimbledon semi-finalist, recomposed herself and eventually powered her way to a 6-3 5-7 6-3 victory.
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The Latvian said: "Of course I could finish it a bit quicker. I got a little bit disappointed after the match point in the second set. But Venus is a great player, a great champion and it's an honour to share the court with her.
"She's a very dangerous player, especially on grass. She was serving really well. It was a little bit hard, but I'm really happy that I managed it and I was fighting until the very last point. She's an idol for a lot of people so it was very special."
Williams, who made her Wimbledon debut in 1997, will now hope to recover in time for another tilt at the All England Club, having been awarded a wild card.
Ostapenko will next play Pole Magdalena Frech, who battled past eighth seed Sorana Cirstea 6-3 6-7 (1-7) 6-4 in a match delayed for nearly two hours by rain.
Britain's Harriet Dart will take on fourth seed Anastasia Potapova, who defeated American Caty McNally on a deciding tie-break.
Meanwhile, there will be an all-Czech quarter-final between top seed Barbora Krejcikova and 18-year-old Linda Fruhvirtova. Krejcikova saw off another Czech player, Tereza Martincova, 6-4 6-4.