Peter Sagan failed in his appeal to be reinstated to the Tour de France a day after he was disqualified for causing a crash.
Sagan was deemed to have elbowed British rider Mark Cavendish into a side barrier during yesterday's fourth stage final sprint in Vittel.
Cavendish broke his shoulder blade in the crash and has had to withdraw from the race.
Meanwhile defending champion Chris Froome has snatched the tour leader's yellow jersey after today's 160km fifth stage, which was won by Italian Fabio Aru.
The Italian champion won the stage after pedalling clear 2.3km from the top of La Planche des Belles Filles, a steep ascent where Britian's Froome himself won in 2012, as did overall race winner Vincenzo Nibali in 2014.
His stage win put Aru third in the overall classification, 14 seconds behind Froome, who took the leader's yellow jersey from Sky team mate Geraint Thomas.
"Froome has shown that he is in excellent condition, much better than on the Dauphine last month where he did not seem to be at his level of the previous years," said the 27-year-old from Sardinia.
"But every time I will have the strength, I will not miss an opportunity to attack him."
Froome was left to regret that the chasing pack had given Aru too much space.
"He attacked just at the right moment, before the last kick," said the three-time Tour de France champion.
Froome, in the words of rival Richie Porte, then 'did his usual squeeze', producing a brutal acceleration that dropped a few riders including Alberto Contador and Nairo Quintana, but mind games between the chasing pack ruined that effort as Aru remained clear.
The Astana rider's team manager Alexandre Vinokourov was delighted with his performance.
"It was an incredible experience to see him like this with the Italian champion jersey on his shoulders, just like Nibali in 2014," said Vinokourov, who was also Nibali's team manager when he won the Tour in 2014. "It gives a lot of confidence."
Aggressive riders who challenged on Wednesday, such as Aru, AG2R La Mondiale's Romain Bardet and Dan Martin of the Quick-Step Floors team, will have another opportunity to unsettle Froome on Sunday.
The 181.5km stage nine route includes three out-of-category climbs before a treacherous descent to the finish at Chambery.