Macclesfield Town's home League Two game against Plymouth has been postponed after the club were served a "zero-capacity notice" at Moss Rose.
The club suspended ticket sales on Friday after being served the notice by the local Safety Advisory Group (SAG).
That led the English Football League to threaten "significant" consequences should Saturday's match be called off.
Macclesfield were docked six points on Thursday for non-payment of wages and failing to fulfil a previous fixture.
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The EFL said that "despite making significant progress, the SAG is not satisfied all its requirements have been met and have maintained the capacity of the stadium at zero".
An earlier statement by Cheshire East Council said the SAG made its decision "against a background of numerous requests to the club to provide the necessary reports and inspection certificates required to demonstrate that the ground is safe".
The EFL claims that the SAG's decision led Macclesfield to inform it that they would not be in a position to fulfil the fixture against Plymouth with supporters in attendance.
However, in a separate statement, Macclesfield's owner Amar Alkadhi claimed the club had "never asked the EFL to play the game behind closed doors".
Alkadhi added that "as far as I am concerned" the game will be played as scheduled.
But in its statement, the EFL insisted the game would not go ahead, while Plymouth's official Twitter account also confirmed the fixture was definitely off.
"The club did request to play behind closed doors, however on consideration, as per the provisions in Regulation 33.1, the EFL did not agree to the request," the EFL said.
"The core principles behind EFL competitions are that all its fixtures are to be played in front of spectators and playing behind closed doors significantly increases the risks of crowds congregating in the vicinity of the stadium.
"As a result of the game not going ahead, the club will be charged with misconduct for failing to fulfil its fixture obligations".
The postponement caps a tumultuous week for Macclesfield after Thursday's six-point deduction, which included an additional four points suspended.
That further deduction could come into effect following another EFL misconduct charge for failing to fulfil fixtures.
A four-point deduction would drop Macclesfield into the League Two relegation zone, as they are currently just three points above it in 22nd place.
The troubled club, who have failed to pay their players on time on numerous occasions in recent months and had a winding-up order over unpaid taxes adjourned for a ninth time on Wednesday after payment was made, must now arrange new dates for matches against Plymouth and Crewe.
Their players refused to play against Crewe on 7 December citing concerns over their emotional wellbeing.