La Liga leaders Real Madrid dropped points at home for only the second time this season when Villarreal fought back from a goal down in an entertaining 1-1 draw that left Real two points ahead of Barcelona on Sunday.
Champions Atletico Madrid also stumbled, a 0-0 stalemate at Sevilla denting their chances of a second consecutive Spanish title and leaving them five points behind Barca in third.
With a King's Cup semifinal second leg against Barca looming on Wednesday, Villarreal coach Marcelino fielded a weakened side at the Bernabeu but it took Real until seven minutes into the second half to break the deadlock.
Eric Bailly hauled Cristiano Ronaldo down in the penalty area and the Portugal captain picked himself up and sent goalkeeper Sergio Asenjo the wrong way with his spot-kick for his 30th goal of the campaign.
Villarreal had the best chances in an even first half. Real keeper Iker Casillas did well to block a close-range Moises Gomez effort in the 12th minute and fullback Dani Carvajal cleared Gerard Moreno's follow-up shot off the line.
Moreno made up for his earlier miss and stunned the European champions when he lashed a shot low into the corner in the 64th minute.
Ronaldo had two chances to restore Real's lead but was twice denied by excellent Asenjo saves before substitute Jese missed an open goal after the Villarreal keeper had misjudged a cross.
The visitors could even have claimed their first win at the Bernabeu when Luciano Vietto, another substitute, tested Casillas with a diving header four minutes from time.
Villarreal withstood intense pressure in the final minutes, Ronaldo again going close with a header.
It was the first game Real have failed to win at home in La Liga since they were beaten 2-1 by Atletico in September and was their first draw in Spain's top flight this season.
Elsewhere
Carlo Ancelotti's side lead on 61 points with 13 games left, with Barca, who host their great rivals at the Nou Camp in three weeks, on 59 after they won a scrappy game at Granada 3-1 on Saturday.
Atletico have 54 points in third, one ahead of Valencia, who won 2-0 at home to Real Sociedad on Sunday and play at Atletico next weekend. Sevilla are seven points further back in fifth.
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