A section of fans in the Southampton end were heard airing the anthem midway through the first half.
The Sex Pistols' version of God Save the Queen was played on the public announcement system at half-time in a mischievous move from the Saints DJ.
Reds fans sparked fury after boos echoed around Wembley Stadium while Prince William - the President of the FA - was introduced to Chelsea and Liverpool players ahead of their cup final clash on Saturday.
Liverpool had the last laugh on Tuesday night as they won 2-1 at St Mary's/
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