- Vincentio – the Duke of Vienna, who also appears disguised as Friar Lodowick
- Angelo – the Duke's deputy who rules in the Duke's absence
- Mariana – Angelo's ex
- Escalus – an ancient lord working under Angelo
- A Justice – friend of Escalus
- The Provost – runs the prison
- Abhorson – the executioner
- Barnardine – a prisoner
- Claudio – a young man imprisoned for impregnating his girlfriend out of wedlock
- Juliet – Claudio's girlfriend, pregnant with their child
- Isabella – Claudio's sister and a novice nun
- Francisca – a nun
- Mistress Overdone – the manager of a brothel
- Pompey – a pimp
- Lucio – a "fantastic"
- Two gentleman – friends of Lucio
- Froth – a foolish gentleman
- Elbow – a simple constable
- Thomas – a friar
- Peter – a friar
- Varrius (silent) – friend of the Duke
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