Maxime Lopez struck for Sassuolo in the 95th minute at the Allianz Stadium on Wednesday to deal Juventus a 2-1 defeat, their first reverse in 10 matches in all competitions.
A stunning finish brought an end to a frustrating encounter for the Bianconeri, whose season has hit another stumbling block after six straight wins heading into Sunday's draw at Inter.
Lopez's breakaway winner decisively put Sassuolo – previously without an away Serie A victory at Juve – in front for the second time, having earlier led through Davide Frattesi.
Weston McKennie's thumping header got Massimiliano Allegri's men back on terms and seemed to set the stage for a second home goal, but it was Sassuolo who were celebrating deep in stoppage time.
The game ended as it began, in breathless fashion. The visitors recovered from a rocky start to go close through Domenico Berardi, whose curling effort tested Mattia Perin, and came through another strong Juventus spell to lead on the stroke of half-time.
Paulo Dybala had hit the post and Federico Chiesa lashed over in the 10 minutes before the break, but an incisive move at the other end gave Frattesi a chance he would not pass up, finishing coolly past Perin.
Giacomo Raspadori was not so accurate when he might have doubled Sassuolo's advantage in the second half, a scare that stirred the hosts.
Juan Cuadrado's blast was cleared off the line by Kaan Ayhan, before Andrea Consigli beat away Dybala's drive and got back to his feet in time to see Chiesa nod over.
McKennie appeared the unlikely saviour then, meeting Dybala's free-kick with a fine header that flew beyond Consigli.
But Juve's pursuit of a second left Lopez in the clear to run onto Berardi's sublime pass, hold off the recovering McKennie and dink over Perin for a memorable Sassuolo triumph.
Coventry City 2-3 Hull City: Tigers win five-goal thriller to keep play-off hopes alive
Hull City kept their hopes of gatecrashing the Championship play-offs alive with a thrilling 3-2 win over beaten FA Cup semi-finalists Coventry City on Wednesday.
Noah Ohio scored the winner with his first touch after coming on as a second-half substitute as Hull moved within three points of sixth-placed Norwich City with two games to play.
Coventry, meanwhile, saw their own hopes of returning to Wembley Stadium for a second successive play-off final all but ended, as they sit six points further back with three matches remaining.
Three days on from their remarkable comeback against Manchester United, the Sky Blues twice fought back to level things at Coventry Building Society Arena. Jaden Philogene's shot squirmed under goalkeeper Bradley Collins to put Hull ahead 31 minutes in, but Kasey Palmer levelled within five minutes with a stunning free-kick.
Hull went back ahead in controversial circumstances on the stroke of half-time, Liverpool loanee Fabio Carvalho converting from the penalty spot despite replays showing Joel Latibeaudiere's foul on Liam Delap took place outside the area.
Mark Robins' men again dragged themselves back into the game as Bobby Thomas headed Jay Dasilva's cross in just before the hour mark, but it was Liam Rosenior's visitors who found a winner.
Thomas went from hero to villain as his poor back pass sold Collins short, and Ohio got a toe to the ball to score the winner, keeping Hull's hopes of Premier League football intact.
Van Dijk: Liverpool have 'no chance' in title race after derby defeat
Virgil van Dijk conceded Liverpool might be out of the Premier League title race after they suffered a surprise 2-0 defeat to Everton in Wednesday's Merseyside derby.
Jarrad Branthwaite and Dominic Calvert-Lewin scored in either half to give the Toffees their first win over Liverpool at Goodison Park since 2010.
Liverpool struggled to find a way through the Toffees' defence despite dominating for long periods, and the Reds' captain says they will not outlast Arsenal and Manchester City without major improvements.
Speaking to Sky Sports, Van Dijk said: "If we play like today, then we have no chance to consider ourselves in the title race.
"We have to just focus on the next game and that will be a tough one anyway, so we will see.
"We're very disappointed, in so many ways. I think everyone has to look at their own performances. Did they really give everything, and do they really want to win the league?
"We're still fighting and there are still games after tonight, but if we play like we did overall in the game today – not winning challenges and giving the referee a chance to give free-kicks like he did many times, then you have no chance to win the title."
Howe: Newcastle performance in Palace defeat 'way off' Spurs win
Eddie Howe admitted Newcastle United's performance in the 2-0 loss at Crystal Palace on Wednesday was "way off" the one that saw them thump Tottenham in their previous game.
Howe's men hammered Spurs 4-0 in their last match, but they never got close to matching the attacking verve from that outing and a Jean-Philippe Mateta double condemned the Magpies to defeat at Selhurst Park.
Newcastle didn't even manage a shot on target until the 87th minute, accumulating just 0.42 xG (expected goals) as the likes of Alexander Isak and Anthony Gordon struggled to make an impact.
Only against Manchester City in August (one) have Newcastle recorded fewer shots on target in a Premier League game this season than their two against Palace, and Howe was critical of his team's display in the loss, one that all but ends their hopes of finishing in the top four.
"[It is a] bad day for us," Howe told Newcastle's club media.
"I thought we were disappointing today, especially in the first 60 minutes. We were way off our levels against Tottenham and our best levels.
"I thought technically we didn't deliver a good performance, and out of possession I don't think it was a particularly good performance either."
Newcastle were unlocked twice for Mateta to score by a Palace side who themselves only generated 0.88 xG.
Howe was left frustrated by the manner in which his side conceded, adding: "I think we'll be disappointed with our performance technically today, and that's probably epitomised by the two goals where we're disappointed with how we've given them the ball in those situations and then how we defended.
"I think the goals sum up aspects of our performance where there was some good, lots of moments that could have led to opportunities, but we were slow to shoot around the box," he said.
The defeat, combined with Manchester United's 4-2 win over Sheffield United, means Newcastle drop to seventh, while eighth-placed West Ham are two points behind with a game in hand.
Newcastle have now lost eight of their last 11 Premier League away games, while they have lost on four occasions in London in the league this term, only doing so more times in a Premier League season in 2000-01 and 2005-06 (both five).
It's been a disappointing season for Howe and his side, especially compared to the heroics of last term when Newcastle finished in the top four, and the Magpies boss knows there's work to be done to get back to those levels.
"It's a day that we have to take our punishment, deal with it, and then try and improve," Howe continued. "It's frustrating because we know we're capable of much better."
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