ACT ONE opens in Windsor castle, where Anne is unhappy, and the courtiers are worried about her future. Anne has failed to bear King Henry a son and his romantic attentions are wandering. He now has his eye on Jane Seymour, one of the queen's ladies. Jane is in love with the king, but she's disturbed by his behavior and concerned about his veiled threats towards Anne.
When Anne's brother, Rochefort, arrives on the scene, he's amazed to see that Lord Percy, Anne's first love, has returned from exile. Percy confides to Rochefort that he's heard rumors of Anne's unhappiness, and that he's been miserable being away from her. Given the king's apparent distrust of Anne, Percy's presence at court seems unwise. But when the king shows up for a hunting party, it turns out that King Henry himself has arranged Percy's return — as a trap for Anne.
If that doesn't put the queen in enough danger, there's also the young man Smeaton, Anne's favorite musician. Smeaton is also in love with the queen, and like many others, he's worried about the king's intentions. Smeaton has a miniature portrait of Anne, which he carries in a locket. He goes to Anne to return it, but has to hide when Rochefort appears.
Rochefort persuades Anne to meet with Percy, who pleads his love. Anne refuses him and Percy draws his sword to kill himself. When Smeaton sees this he thinks Anne is in danger, and rushes out from his hiding place. At the same moment, the king suddenly appears.
Smeaton declares that Anne has done nothing wrong, but then he accidentally drops her portrait at the king's feet. The king orders his guards to detain everyone. Anne begins a spectacular sextet by trying to explain the situation. But as the act ends the king says she'll have to defend herself in court, and Anne knows there may be no way to save herself.

In ACT TWO, Jane tells Anne that the king will spare her if she confesses to her crimes. Jane also reveals that King Henry loves another woman. Anne demands to know who it is, and the confrontation leads to a powerful duet. Torn by her love for Henry and her loyalty to Anne, Jane confesses the she is the king's other woman.
At Anne's trial, Smeaton lies and says he is Anne's lover. He's been told that his false confession will save Anne's life — but instead it assures her death. Percy is willing to die to save Anne, and Jane also pleads with the king to let Anne live. But King Henry is determined to see things through.
Alone in the Tower of London, Anne tenderly recalls her childhood home and her early love for Percy. As she begins to pray, bells announce the king's marriage to Jane. To music combining desperation with ecstasy, Anne asks for God's mercy on the king and his new wife. Then she's led away to the execution block as the opera ends.
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