Fig. 6

ω-3 PUFA supplementation enhances autophagy via promoting nuclear export of Beclin-1 in lesioned cortices 7 days after TBI. a, b Immunofluorescence staining and western blot analyses demonstrated that expression levels of Beclin-1 in the cytosol, nuclei, and in total protein from lesioned cortices increased 7 days after TBI and that ω-3 PUFA supplementation effectively increased Beclin-1 expression in the cytosol and in total protein of cells from lesioned cortices (p < 0.05), but not in nuclear protein (p > 0.05). Moreover, less cytoplasmic redistribution of nuclear Beclin-1 was found in the presence of an autophagy inhibitor, 3-MA, after TBI. Representative photomicrographs of Beclin-1-positive neurons are shown (× 400). c Co-IP assays confirmed that ω-3 PUFA supplementation significantly increased interactions between cytoplasmic Beclin-1 and Bcl-2 after the TBI, while 3-MA treatment reversed these increases. Values are expressed as mean ± standard deviation (n = 6 per group). N.S., p > 0.05, *p < 0.05, **p < 0.01. Scale bars = 50 μm