Karl V, Holy Roman Emperor (b. 1500) married a) Mary Tudor (1496-1515) in 1514, b) Anne of Hungary (b. 1503) in 1516
- a) Philip I, Holy Roman Emperor (b. 1515) married Sybille, Duchess of Jülich-Cleves-Berg (b. 1512)
- b) Eleanor (b. 1521)
- b) Maximilian, King of Hungary and Bohemia (b. 1522)
- b) Elisabeth (b. 1523)
- b) Anna (b. 1525)
- b) Margaret (b. 1526)
- b) Frederick (1527)
- b) Joanna (1529-1532)
- b) Magdalena (b. 1530)
- b) Agnes (b. 1531)
- b) Charles (b. 1533)
- b) Helena (b. 1536)
Isabella of Austria (b. 1501) married a) Christian II, King of Denmark (1481-1520) in 1514 b) Antoine, Duke of Loraine (b. 1489)
- a) John II, King of Denmark, Sweden and Norway (b. 1518)
- a) Philip Ferdinand (1519), twin of Maximilian
- a) Maximilian (1519), twin of Philip Ferdinand
- a) Dorothea of Denmark (b. 1520)
- b) Anne of Lorraine (b. 1522)
- b) Charles III, Duke of Lorraine (b. 1524)
- b) Isabella of Lorraine (b. 1528)
- b) Eleanor of Lorraine (b. 1530)
Eleanor of Austria (b. 1498) married Francis I, King of France (b. 1494) in 1517
- Charles IX, King of France (b. 1518)
- Isabella (b. 1520)
- Francis, Duke d’Orleans (1522-1530)
- Madeleine (b. 1525)
- Margaret (1526)
- Eleanor (b. 1528) married Francis III, Duke of Brittany (b. 1531)
Mary/Margaret ? (b. 1505) married a) Louis of Hungary and Bohemia (1506-1515) in 1515, b) Sigismund I, King of Poland (b. 1467) in 1519
- b) Sigismund II of Poland (b. 1522)
- b) Eleanor (b. 1524)
- b) Isabella (b. 1526)
- b) Albert (1528)
- b) miscarriage (1529)
- b) stillborn son (1531)
- b) Margaret (1532)
Claude, Duchess of Brittany (1499-1516) married Francis I, King of France (b. 1494)
- Louise, Duchess of Brittany (1515-1517)
- Charlotte, Duchess of Brittany (b. 1516-1527)
James IV, King of Scotland (b. 1512) married Renée, Duchess of Brittany (b. 1510) in 1526
- Anne (b. 1528) married Alexander Stewart, Duke of Albany and Count of Auvergne and Boulogne (b. 1519) *
- James VI, King of Scotland (b. 1530) married
- Francis III, Duke of Brittany (b. 1531) married Eleanor of France (b. 1528)
- Margaret (1532-1535)
*son of John of Albany and his cousin and second wife Madeleine de la Tour d‘Auvergne (who he married after Anne’s death in 1515)