Atletico Madrid suffered a blow in the race for a top-three finish in La Liga as Roberto Soriano's late goal gave Villarreal a 1-0 win at the Vicente Calderon.
Diego Simeone's side were looking to keep themselves clear ahead of Sevilla in the battle for Champions League qualification.
However, Los Colchoneros could not make the most of their possession and were made to pay when Italy forward Soriano converted a cross from fellow substitute Cedric Bakambu with eight minutes left.
Villarreal, in fifth place, close up to five points behind Sevilla, who can go level with Atletico should they beat Celta Vigo at home on Thursday.
Visiting goalkeeper Andres Fernandez saved from France forward Antoine Griezmann and midfielder Saul Niguez before also thwarting Nicolas Gaitan in a one-on-one during the second half.
Atletico's frustrating evening was compounded by the loss of Belgium playmaker Yannick Carrasco with what appeared to be a shoulder injury not long after he was sent on as a second-half substitute -- leaving Simeone with plenty to ponder as the Champions League semifinal against city rivals Real Madrid looms.