İmamoğlu's arrest and imprisonment have been widely viewed as politically motivated despite Turkey's government insisting the judiciary is independent and free from political influence.
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Istanbul's jailed opposition mayor has appeared in court in one of multiple cases against him, prompting hundreds of supporters to gather outside the prison where the hearing was taking place.
Ekrem İmamoğlu has been held at Silviri Prison west of Istanbul since his formal arrest on corruption charges on 23 March.
The current court case alleges that he threatened a public prosecutor, and is one of six that predate his arrest last month, sparking nationwide protests.
Addressing the judge, İmamoğlu said he was in court because he had won three elections against the person "who thinks he owns Istanbul," a reference to President Recep Tayyip Erdoğan, who launched his political career as the city's mayor in the 1990s.
Erdoğan was heavily involved in backing his party's candidates against İmamoğlu.
The hearing was attended by İmamoğlu's wife and son, as well as lawmakers from the Republican People's Party (CHP), Halk TV and other media outlets.
The case was adjourned until 16 June.
The mayor, who is also the main opposition challenger to Erdoğan's 22-year rule at the next presidential election, faces more than seven years in prison and a political ban for allegedly "targeting, threatening and insulting persons working in the fight against terrorism."
The charge stems from comments he made on 20 January in which he criticised Istanbul Chief Public Prosecutor Akın Gürlek over criminal cases brought against other opposition figures.
İmamoğlu was initially detained on 19 March in relation to two investigations, one focusing on corruption in the Istanbul municipality and another alleging terrorism links in his party's electoral pact with pro-Kurdish politicians.
Demonstrations calling for his release and an end to Turkey's democratic backsliding under Erdoğan have led to the arrest of around 2,000 people for attending rallies banned by city authorities.
The mayor was officially nominated as the CHP presidential candidate while in custody.
An election is due to be held in 2028 but may come sooner and İmamoğlu's imprisonment has been widely viewed as politically motivated despite Turkey's government insisting the judiciary is independent and free from political influence.
Also on Friday, two other courts in Istanbul were also holding hearings on cases against İmamoğlu.
One is a bid-rigging case that dates back 10 years, when he was mayor of Istanbul's Beylikdüzü district.
The other alleges illegal donation collection and stems from a video circulated in the run-up to last year’s local elections showing CHP staff counting bundles of cash.