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. 2017 Aug 14;12(8):e0183076. doi: 10.1371/journal.pone.0183076

Fig 5. Effects of PINK1 G309D mutant on the signaling pathways involved in MG132-induced HO-1 expression and Nrf2 activation in SH-SY5Y cells.

Fig 5

(A) SH-SY5Y cells were pretreated for 30 min with LY294002 (20 μM), PD98059 (20 μM), SB23580 (10 μM) or N-Acetyl-L-cysteine (NAC) (10 mM). Subsequently, MG132 (1 μM) was added for 24 h. (B) SH-SY5Y cells were were pre-treated for 30 min with LY294002 (20 μM) or SB23580 (10 μM). Then, MG132 (1 μM) was added for 6 h, and the nuclear extracts were then prepared for the evaluation of Nrf2 protein levels. Note that PI3K/Akt inhibitor, p38 MAPK inhibitor, and NAC significantly antagonized the induction of HO-1 by MG132. Furthermore, PI3K/Akt or p38 MAPK inhibitor inhibited MG132-induced Nrf2 nuclear translocation. MG132 time-dependently increased p38 (C) and Akt (D) phosphorylation and expression of recombinant PINK1 G309D mutant inhibited the phosphorylation of p38 and Akt following MG132 treatment. Results are expressed as multiples of controls (mean ± S.E.M) (n = 3). *p < 0.05 compared with control group (con.); #p < 0.05 compared with MG132 treatment alone.