Almost exactly one year after opposition figure Leopoldo Lopez was arrested as he led a wave of protests against President Nicolas Maduro, intelligence agents burst into Mayor Antonio Ledezma's office late Thursday and hauled him to jail.
A third radical critic of the government, ousted lawmaker Maria Machado, is under investigation over an alleged plan to assassinate Maduro, though she remains free.
Ledezma, 59, will be held in Ramo Verde prison, the same jail on the outskirts of the capital currently housing Lopez, officials said in ordering his continued detention.
Opposition leader Henrique Capriles called on the government to produce evidence of the supposed conspiracy.
"Does Maduro think that putting everyone in prison is going to get him 50 popularity points or that he's going to win elections?" the two-time presidential candidate asked.
Maduro, who has accused the opposition of trying to topple him several times since his April 2013 election, said late Thursday that the mayor was detained over a coup plot financed by the United States.
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