Fig. 4. Inhibition of mitophagy decreased the protective effects of NR on H₂O₂-treated 661w cells.

a Inhibition of NR-induced increase of co-localized puncta (LC3 and TOM20) after 1 μmol/L CsA pretreatment. b, c Western blot analysis of LC3 and p62. The relative expression of LC3B II and p62 were normalized to β-actin (n ≥ 3). d, f Cell viability in the CQ + NR + H₂O₂ and CsA +NR + H₂O₂ treatments was lower than that of the NR + H₂O₂ group (n = 5). e 1 μmol/L CsA pretreatment inhibited the NR-induced increase of mitochondrial membrane potential in H₂O₂-treated 661w cells (n = 6). g 50 μmol/L CQ pretreatment inhibited the NR-induced decline of ROS generation in H₂O₂-treated 661w cells (n = 5). Data are shown as mean. Scale bar: 25 μm.