Gian Francesco II. Gonzaga (1466-1519), Margrave of Mantua

Isabella d'Este (1474-1539)

1. Maria (c. 1509-1530), first daughter of Margrave Guglielmo IX. del Montferrat († 1518); marriage in 1516; annulment of the marriage by Pope Clement VII. in 1528
2. Julia of Aragon (1492-1542), daughter of King Federigo of Naples († 1504) and Isabella del Balzo († 1533); marriage on 9. April 1531; annulment of the marriage by Pope Clement VII. in March 1532
3. Margherita (1510-1566), daughter of Margrave Guglielmo IX. del Montferrat († 1518); marriage on 3. October 1532

1. Francesco III., Duke of Mantua; born on 10. March 1534, deceased on 22. February 1550 (drowned); married to Katharina of Austria (1533-1572), daughter of Emperor Ferdinand I. († 1564)
2. Isabella, born on 18. April 1536/37, deceased on 16. August 1579; married to Don Ferrante Francesco d'Avalos d'Aquino of Aragon, Margrave of Pescara
3. Guglielmo I. (or William I.), Duke of Mantua; born on 24. April 1538, deceased on 14. August 1587; married to Eleonore of Austria (1534-1594), daughter of Emperor Ferdinand I.; his son Vincenzo I. (1562-1612) became Duke of Mantua, he was married to Eleonora de' Medici, the eldest daughter of Francesco I. de' Medici († 1587)
4. Luigi, Duke of Nevers; born on 18. September 1539, deceased on 23. October 1595
5. Federigo, Cardinal; born in 1540, deceased on 21. February 1565