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| Charles I |
| Children |
| Felipe, Prince of Asturias (future Philip II) |
| Infanta Maria, Holy Roman Empress |
| Infanta Juana, Crown Princess of Portugal |
| Infante Juan (natural son) |
| Infanta Margarita, Duchess of Florence, Duchess of Parma (natural daughter) |
| Philip II (I of Portugal) |
| Children include |
| Carlos, Prince of Asturias |
| Infanta Isabella, Archduchess of Austria |
| Infanta Catalina, Archduchess of Austria, Duchess of Savoy |
| Felipe, Prince of Asturias (future Philip III) |
| Philip III (II of Portugal) |
| Children include |
| Infanta Ana, Queen of France |
| Felipe, Prince of Asturias (future Philip IV) |
| Infanta Maria Ana, Holy Roman Empress |
| Infante Carlos |
| Cardinal-Infante Fernando |
| Philip IV (III of Portugal) |
| Children include |
| Baltasar Carlos, Prince of Asturias |
| Infanta Maria Theresa, Queen of France |
| Infanta Margarita, Holy Roman Empress |
| Infante Carlos (future Charles II) |
| Charles II |
