Wooden breast myopathy (WB) is characterized by palpation of a rigid pectoralis major (p. major) muscle and is under severe oxidative stress and inflammation. Previous studies have shown that vitamin E (VE) has antioxidant property and omega-3 (n-3) fatty acids have an anti-inflammatory effect. The objectives of this study were to identify the effects of VE and n-3 fatty acids on the severity of WB, morphological structure of p. major muscle, and expression of gene associated with WB, and to determine the most beneficial supplementation period. A total of 210 Ross 708 broilers were randomly assigned into seven treatments with 10 replicates of three birds each. The control group was fed with corn-soybean meal basal diet during the entire study (0 to 58 d). Supplementation of VE (200 IU/kg), n-3 fatty acids (n-6/n-3 ratio of 3:1), or combination of both was performed during starter phase (0 to 10 d) or grower phase (11 to 24 d). All of the broilers were harvested at 58 d. Muscle fiber width, perimysial and endomysial connective tissue space, morphology and WB scores were obtained. Gene expression level was measured through Nanostring analysis. Microscopic evaluation showed that supplemental VE in the grower diet increased percentage of muscle with no WB to 82%. Genes associated with muscle growth and inflammation were differentially expressed in the group supplemented with VE in grower diet. These data suggest that supplementation of VE in grower diet may reduce the severity of WB which will benefit the broiler industry.