Germany's players covered their mouths for their team photo before their opening World Cup match, a 2-1 loss to Japan, to protest against FIFA's clampdown on the 'One Love' armband.
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The Germany team lined up in the traditional formation before the game and all 11 players covered their mouths with their right hand in a co-ordinated gesture.
"It was a sign from the team, from us, that FIFA is muzzling us," Germany coach Hansi Flick said after his side's shock defeat.
The gesture was a response to the world governing body's effective nixing of seven European teams' plans to wear armbands that were seen as a rebuke to host nation Qatar and its human rights record.
Team captains, including Germany captain Manuel Neuer, had planned to wear the armband - which features a heart-shaped, multicoloured logo, and is a symbol for inclusion and diversity.
But the seven federations - which also include England, Wales, Belgium, the Netherlands, Switzerland and Denmark - backed down after FIFA made it clear they would face consequences if they did not.
"We may have our bands taken away from us, but we'll never let our voices be taken from us," Neuer said.
"We stand for human rights. That's what we wanted to show. We may have been silenced by FIFA regarding the captain's armbands, but we always stand for our values."
FIFA said Germany will not face disciplinary action for the gesture.
The German soccer federation posted a statement on Twitter during Wednesday's game.
"With our captain's armband, we wanted to send a signal for values that we live in the national team: diversity and mutual respect. Being loud together with other nations," they wrote.
"It's not a political message: human rights are nonnegotiable. That should be obvious. Unfortunately it still isn't. That's why this message is so important to us.
"Denying us the armband is like muzzling us. Our stance stands."
Qatar has been under scrutiny for its human rights record and laws criminalising homosexuality.
FIFA issued its warning to the federations on Monday, hours before England and the Netherlands were set to play with their captains wearing the One Love armbands.
The governing body said players would immediately be shown a yellow card and could face further consequences.
German Interior Minister Nancy Faeser wore a One Love armband in the stands for her country's match, sitting beside FIFA's president Gianni Infantino.
Faeser posted a photo of herself at the game with the hashtag #OneLove.
Belgium's foreign minister also sported the armband when she met with Infantino during her country's 1-0 win against Canada on Wednesday (Thursday AEDT).
Hadja Lahbib was pictured wearing the colourful item on her left arm as she spoke with Infantino, and subsequently posted a photo of herself on Twitter.
Australian Associated Press