In addition to conflicts between his Spanish and German inheritances, conflicts of religion would be another source of tension during the reign of Charles V. Before Charles's reign in the Holy Roman Empire began, in 1517, Martin Luther launched what would later be known as the Reformation. At this time, many local dukes saw it as a chance to oppose the hegemony of Emperor Charles V. The empire then became fatally divided along religious lines, with the north, the east, and many of the major cities – Strasbourg, Frankfurt, and Nuremberg – becoming Protestant while the southern and western regions largely remained Catholic.
The record of an October 1517 visit by Louis d'Aragon states that the Mona Lisa was executed for the deceased Giuliano de' Medici, Leonardo's steward at the Belvedere Palace between 1513 and 1516—but this was likely an error. According to Vasari, the painting was created for the model's husband, Francesco del Giocondo. A number of experts have argued that Leonardo made two versions (because of the uncertainty concerning its dating and commissioner, as well as its fate following Leonardo's death in 1519, and the difference of details in Raphael's sketch—which may be explained by the possibility that he made the sketch from memory). The hypothetical first portrait, displaying prominent columns, would have been commissioned by Giocondo circa 1503, and left unfinished in Leonardo's pupil and assistant Salaì's possession until his death in 1524. The second, commissioned by Giuliano de' Medici circa 1513, would have been sold by Salaì to Francis I in 1518 and is the one in the Louvre today. Other believe that there was only one true Mona Lisa, but are divided as to the two aforementioned fates. The famed painting was kept at the Palace of Fontainebleau, where it was kept until Louis XIV moved it to the Palace of Versailles, where it remained until the French Revolution. In 1797, it went on permanent display at the Louvre.