ABSTRACT: The Laetiporus sp. is a fungal species that is traditionally used for medicinal purposes. This study investigated the effects of the Laetiporus sulphureus fermented product (FL) as a feed supplementation on the antioxidant activities, the intestinal Tight Junction (TJ) mRNA expression, and the intestinal morphology of broiler chickens. Four-hundred one-day-old male broilers (Ross 308) were randomly allocated to five experimental diets: (1) a corn-soybean meal basal diet (control), (2) a basal diet replaced with 5% Wheat Bran (5% WB), (3) a basal diet replaced with 10% WB (10% WB), (4) a basal diet replaced with 5% FL (5% FL), and (5) a basal diet replaced with 10% FL (10% FL). The FL-supplemented groups exhibited a better feed conversion ratio in the overall experimental period compared to the WB and control groups. The serum antioxidant profiles of 35-day-old broilers showed that, compared to the control and 10% WB groups, the 5% FL supplementation group had a significantly increased superoxide dismutase activity, while it down-regulated the concentration of malondialdehyde in the serum (p < 0.05). The assessment of selected antioxidant gene expression showed that the 5% FL group significantly elevated heme oxygenase-1 and nuclear factor erythroid 2-related factor 2 expression, compared to the control and WB groups (p < 0.05). Furthermore, both of the FL supplemented groups had a significantly higher expression of glutathione peroxidase and catalase, compared to that of the WB and control groups in the jejunum (p < 0.05). The TJ mRNA expression in the jejunum showed that 5% FL significantly elevated the zonula occludens-1, claudin-1, and mucin-2 expression (p < 0.05), while 5% and 10% FL supplementation significantly improved OCLN expression in both the jejunum and ileum, compared to control group (p < 0.05). The intestinal morphology of 35-day-old broilers showed that a 5% FL supplementation significantly increased the villus height in the ileum and jejunum, compared to the WB and control groups (p < 0.05). Moreover, the 5% and 10% FL supplementation groups had a significantly higher villi:crypt ratio in the ileum, compared to the WB and control groups (p < 0.05). To conclude, FL supplementation improved the antioxidative status, the TJ mRNA expression, and the intestinal morphology, and it was accompanied by a lowered feed conversion ratio in broilers. Finally, 5% supplementation had the overall best results in improving the antioxidant status, TJ mRNA expression, and intestinal morphology of broilers.