In 1584 Gentili and Jean Hotman (1552—1636) were asked by the government to advise on the treatment of Spanish ambassador Bernardino de Mendoza (about 1540–1604), who had been implicated in the so-called Throckmorton plot against Queen Elizabeth I.
He was expelled from England in 1584 after his involvement in Francis Throckmorton's plot against Elizabeth I was revealed.
Following his return to England in 1583, he served as an intermediary for communications between supporters of the Catholic cause on the continent, the imprisoned Mary, Queen of Scots, and the Spanish ambassador Bernardino de Mendoza.
With Lord Henry Howard and Francis Throckmorton he was arrested on suspicion of complicity late in the same year, and for a second time was sent to the Tower.
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