Fig. 4.

ω-3 PUFA supplementation promotes autophagic flux on rat hippocampal neurons in vitro. a Autophagy markers LC3 with GFP and RFP protein which indicate real-time autophagy flux levels were imaged by confocal microscope (× 400). Representative photomicrographs of autophagy flux in neurons are shown. b Bar graphs displayed the mean ± standard deviation of the LC3 puncta per cell, which indicated that the values of the ω-3 related groups were significantly different from those of the control group. The control group maintained basal levels of autophagy. The ω-3 PUFA group showed a suppression of autophagy at late stages compared to the early stage suppression of the control group, while SIRT1 siRNA or autophagy inhibitor reversed ω-3 PUFA-mediated increases in autophagy. c Western blot showed a significant increased LC3 expression and decreased p62 levels in the ω-3 PUFA treatment group. However, the SIRT1 siRNA or autophagy inhibitor groups showed an increase in p62 levels but no increases in LC3 expression. Values are expressed as mean ± standard deviation (n = 6 per group). N.S., p > 0.05, *p < 0.05, **p < 0.01