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. 2020 Jul 30;11(7):607. doi: 10.1038/s41419-020-02750-0

Fig. 6. ω-6 PUFAs increase autophagy levels via activation of the antioxidant pathway.

Fig. 6

Representative confocal microscopic image of hepatocytes stained with LysoTracker after treatment with a Nrf2 pathway inhibitor (5 μM ML385) or activator (2 μM NK-252). Scale bars, 50 μm (a, c). The presence of LysoTracker-stained intracellular autolysosomes was demonstrated by flow cytometry, and the numbers of the autolysosomes were calculated by flow cytometric analysis of the mean red (FL4) fluorescence intensity after treatment with a Nrf2 pathway inhibitor or activator (b, d). Nuclear Nrf2 was examined by western blot analysis and quantitated after treatment with a Nrf2 pathway inhibitor (n = 3) (e, g). The mRNA expression levels of key antioxidation-related genes (Nrf2, Sod1, Sod3, CAT, and Gpx) were analyzed after treatment with a Nrf2 pathway inhibitor or activator (n = 6) (f, h). The results are presented as the mean with SEM and were analyzed using Tukey’s test. Bars bearing the same letters are not significantly different among treatments (*P ≥ 0.05).