Almost two thirds of people who voted in South Yorkshire opted to leave the European Union, backing the national outcome.
Aside from a narrow leave vote in Sheffield, where 49% opposed Brexit, the Leave campaign saw huge support in Barnsley, Doncaster and Rotherham.
UKIP leader Nigel Farage hailed the result in Sheffield as "amazing".
In total, 61.56% of people in the region voted to leave and 38.44% voted to remain in the EU.
Across the Yorkshire and Humber region, 58% of voters backed leave while nationally the UK voted to Leave by 52% to 48%.
Amjad Bashir, Conservative MEP for Yorkshire and the Humber and Leave campaigner said people had "nothing to be worried about" from the decision to leave the EU and that it was time to "look forward to a bright new future".
Labour MP for Penistone & Stocksbridge Angela Smith has called for Labour Party leader Jeremy Corbyn to consider his position after failing to mobilise the Party's core support.
She said: "Given the challenges we face as a country we've got to have a decent, robust leadership in the Labour party that puts the country first and on that basis I'm asking Jeremy to really consider his position and to consider whether or not he's capable of meeting that challenge."
BBC Radio Sheffield listener reaction
- "Best news ever today about getting away from the corrupt EU. It's time to get our country back and away from the soft, lefty, Labour voters who would see us give everything we've worked for away to any number of work-shy scroungers"
- "I decided last night I was voting out because I wanted to give the government a bloody nose. I never expected the Leave vote would carry. I felt like I had been nasty to somebody and done something to them and then instantly regretted it"
- "The world won't end, but for the last 30 years South Yorkshire has rebuilt itself on EU investment - just wonder if we will be forgotten by central government yet again"
Wentworth and Dearne MP John Healey said the result in South Yorkshire was a "big disappointment".
Asked why Labour had failed to get its Remain message through in one of the party's traditional strongholds, he said: "Labour voters like the rest of the country are divided about this question."
Analysis, BBC Look North political editor Len Tingle
This was the campaign where the Remain camp underestimated the simple clear message being voiced by Boris Johnson and Nigel Farage on their frequent barn-storming campaign tours through Yorkshire.
They repeatedly voiced anger at the "big brother" EU spending our money in ways which are not in our best interests. It struck a chord with voters even in areas like South and West Yorkshire where over the years billions of Euros have been spent on regional reconstruction packages.
In the agricultural market towns and fishing ports, latent euro scepticism was deeply enhanced. The prime minister and the chancellor were simply disbelieved and Labour leader Jeremy Corbyn's message was received as being so half-hearted as to be worthless.
Doncaster MP Ed Miliband, on the last day of campaigning in Leeds, made a plea to voters not to use this as an opportunity to punish the government or "the establishment".
Looking at the overwhelming voter support in Yorkshire to quit the EU, his message too fell on largely deaf ears.
Liberal Democrat peer and former Sheffield Council leader Lord Scriven told BBC Radio Sheffield the result had filled him with "absolute despondency and despair for future generations".
Steve Winstone, UKIP representative for Sheffield, said he was "shocked that the people have spoken in such a way" but that the result was "fantastic".