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Katharine Hepburn
Mary Stuart
Florence Eldridge
Elizabeth Tudor
Fredric March
Bothwell
Douglas Walton
Darnley
John Carradine
Rizzio
Robert Barrat
Morton
Gavin Muir
Leicester
Ian Keith
Moray
Moroni Olsen
John Knox
William Stack
Ruthven
Ralph Forbes
Randolph
Alan Mowbray
Throckmorton
Frieda Inescort
Mary Beaton
Donald Crisp
Huntly
David Torrence
Lindsay
Molly Lamont
Mary Livingstone
Anita Colby
Mary Fleming
Jean Fenwick
Mary Seton
Lionel Pape
Burghley
Alec Craig
Donal
Mary Gordon
Nurse
Monte Blue
Messenger
Robert Warwick
Sir Francis Knollys
Halliwell Hobbes
Man (uncredited)
Nigel De Brulier
Judge