Stevenage have been relegated to League Two following their 3-1 defeat to Bristol City.
Graham Westley's side knew going into their game at the Lamex Stadium that only a win could see them hang onto their place in League One.
They fell behind in the 42nd minute to a Wade Elliott strike, and after the break Sam Baldock and Stephen Pearson put City 3-0 ahead to put the final nails in the Stevenage coffin.
Luke Freeman scored a late consolation goal, but the club's relegation was confirmed at the final whistle.
Peterborough United took a huge stride in securing the final playoff place in League One as they hammered Carlisle United 4-1 at London Road.
Conor Washington put the Posh ahead after 14 minutes while Britt Assombalonga added a second from the penalty spot.
Lee Miller did get a goal back for the visitors in the 63rd minute, but Assombalonga fired in another spot kick and Michael Bostwick made sure of the win with the fourth in the dying minutes.
Swindon Town kept up the pressure on Peterborough though as they beat Bradford City 1-0 in their clash at the County Ground.
Lee Cox scored the only goal of the game in the 64th minute to leave the Robins four points off sixth place with two games remaining in the season.
Notts County kept their survival hopes alive as they recorded a narrow win over Crawley Town. James Spencer's strike was enough to hand them all three points to move them within a point of safety.
Shrewsbury Town are all but down as they fell to a heavy 5-2 defeat to Preston North End at Deepdale.
Paul Gallagher scored a second-half hat-trick for Simon Grayson's side to hand them the win along with strikes from Lee Holmes and Bailey Wright, while Shaun Miller and Luke Summerfield netted for the Shrews.
Rotherham United strengthened their hold on third place in the table as a Kieran Agard goal handed them a 1-0 win over Port Vale, despite playing the second half with 10 men after Richard Smallwood was sent off.
Cody McDonald snatched a point for Gillingham in their clash with Walsall at Bank's Stadium after Febian Brandy had handed the home side the lead.
Meanwhile, the games between Tranmere Rovers and Sheffield United, and Oldham Athletic and Coventry City both ended goalless.
Results in full: Crewe Alexandra 0-0 Colchester United, Leyton Orient 1-3 Wolverhampton Wanderers, MK Dons 2-2 Brentford, Notts County 1-0 Crawley Town, Oldham Athletic 0-0 Coventry City, Peterborough United 4-1 Carlisle United, Preston North End 5-2 Shrewsbury Town, Rotherham United 1-0 Port Vale, Stevenage 1-3 Bristol City, Swindon Town 1-0 Bradford City, Tranmere Rovers 0-0 Sheffield United, Walsall 1-1 Gillingham.
