- Saoirse Ronan María Estuardo
- Margot Robbie Isabel I
- Jack Lowden Lord Darnley
- Joe Alwyn Robert Dudley
- David Tennant John Knox
- Guy Pearce Sir William Cecil
- Gemma Chan Bess de Hardwick
- Martin Compston Conde de Bothwell
- Brendan Coyle Conde de Lennox
- Ian Hart Lord Maitland
- Adrian Lester Lord Randolph
- Ismael Cruz Cordova David Rizzio
- James McArdle Conde de Moray
- Maria Dragus Mary Fleming
- Izuka Hoyle Mary Seton
- Eileen O'Higgins Mary Beaton
- Liah O'Prey Mary Livingston