Pasture- and feedlot-finished beef steers (n = 48) were supplemented with 25-hydroxyvitamin D3 (25-OH D3) on d 7 prior to harvest. The longissimus (LM), semimembranosus (SM), and gracilis (GR) muscles were collected after harvest for evaluation of effects on sensory traits and fresh beef color. Sensory analysis indicated that supplementation of 25-OH D3 improved tenderness of GR steaks in pasture finished steers, but not feedlot-finished steers. Tenderness of the SM and LM were not affected by 25-OH D3 supplementation. This seems to indicate that muscles are differentially affected by 25-OH D3. Furthermore, as aging time increased, L* values increased, indicating steaks were lighter with increased aging time. Also, results indicated that pasture-finished steers yield steaks that are less red than steaks from feedlot-finished steers. This may have implications for consumers as they generally prefer a bright, cherry-red colored product.