Charles V, Holy Roman Emperor (b.1500: d.1558) m. Katherine Tudor (b.1503: d.1531) (a), Louise of France (b.1515: d.1559) (b)
1a) Isabella of Austria (b.1521: d.1524)
2a) Charles II of Spain (b.1523: d.1570) m. Christina of Denmark (b.1521: d.1590) (a)
1a) Maria of Spain (b.1544)
2a) Alfonso of Spain (b.1546: d.1548)
3a) Charles III of Spain (b.1547)
4a) Isabella of Spain (b.1550)
3a) Ferdinand of Austria, Duke of Burgundy (b.1524: d.1558) m. Sophia Jagiellon (b.1522: d.1575) (a)
1a) Philip V, Duke of Burgundy (b.1547)
2a) John of Burgundy (b.1550: d.1550)
3a) Elisabeth of Burgundy (b.1552)
4a) Philip of Austria (b.1526: d.1528)
5a) Maria of Austria (b.1528: d.1600) m. Maximilian, King of Hungary, Bohemia and Croatia (b.1527: d.1576) (a)
1a) Anna of Hungary (b.1546)
2a) Maria of Hungary (b.1547)
3a) Ferdinand II of Hungary, Bohemia and Croatia (b.1549)
4a) Elisabeth of Hungary (b.1552)
5a) Matthias of Hungary (b.1555: d.1556)
6a) Margaret of Hungary (b.1558)
6a) Philip of Austria (b.1530: d.1545)
7b) Joanna of Austria (b.1533: d.1599) m. Alfonso II d'Este, Duke of Ferrara (b.1533: d.1597) (a)
8b) Margaret of Austria (b.1534: d.1601) m. João Manuel, Prince of Portugal (b.1537: d.1554) (a)
1a) John IV of Portugal (b.1553)
2a) Catherine of Portugal (b.1554)
9b) John of Austria, Duke of Milan (b.1537: d.1609) m. Eleonora Gonzaga (b.1531: 1599) (a)
1a) Isabella of Milan (b.1551)
2a) Madeleine of Milan (b.1553)
3a) Federico of Milan (b.1555: d.1556)
4a) John of Milan (b.1558)
10b) Stillborn Boy (c.1540)
11b) Stillborn Boy (c.1541)
12b) Catherine of Austria (b.1543: d.1555)
13b) Stillborn Girl (c.1545)
14b) Stillborn Boy (c.1546)
15b) Claudia of Austria (b.1548: d.1613) m. Charles IX of France (b.1550: d.1574) (a)
1a) Marie Elisabeth of France (b.1569)
2a) Charles X of France (b.1571)
3a) Claude of France (b.1572)
4a) Louis, Duke of Anjou (b.1574)
5a) Elisabeth of France (b.1575: d.1575)