Elisabeth was born in the Château de Fontainebleau. She was raised under the supervision of the governor and governess of the royal children, Jean d'Humières and Françoise d'Humières.
Elisabeth's childhood was spent in the French royal nursery, where her father insisted she share her bedroom with her future sister-in-law, Mary, Queen of Scots, who was about three years older. Although Elisabeth had to give precedence to Mary (since Mary was already a crowned queen), the two would remain close friends for the rest of their lives.
Her lady-in-waiting, Claude de Vineulx, accompanied her to Spain and often wrote reports of Elisabeth's health to Catherine. She was described as being shy, timid and very much in awe of her formidable mother, even as there is evidence in letters that Catherine was tender and loving towards her. While it is acknowledged that her sister Margaret and her future sister-in-law Mary were both prettier than she, Elisabeth was nevertheless still considered one of Catherine's attractive daughters.

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