Born in Kansas City, Missouri, Truex learned acting at an early age after his father, a doctor, treated actor Edwin Melvin, who paid his bill by giving the son elocution lessons. He started acting at age five and toured through Missouri at age nine as "The Child Wonder in Scenes from Shakespeare".
As a young man, he lived in Denver and was among the supporting actors at the Elitch Theatre, appearing during the 1903,1904,1905,1906 and 1907 seasons. Ernest Truex began his career of "walk-ons" at Elitch while he was still a student at East High School (where his classmates included Douglas Fairbanks and Harold Lloyd.) Among his performances at Elitch Theatre were the play of When Knighthood Was in Flower with Maude Fealy. and Tess of the d'Urbervilles, which featured Tyrone Power Sr. and a young Cecil B. DeMille in the supporting cast. In 1906, he appeared in several shows with young Colorado natives, Douglas Fairbanks and Spring Byington.
His Broadway debut came in Wildfire (1908), and he performed in several David Belasco plays and portrayed the title role in the 1915 musical Very Good Eddie. Truex played the lead role in the disastrous 1923 premiere of F. Scott Fitzgerald's The Vegetable. In 1927, he created the role of Bill Paradene in Good Morning, Bill, which was based on an original play by Ladislas Fodor and adapted by P.G. Wodehouse.

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