Hibitor, PD98059 (Figure 6A), suggesting a vital part sulfuretininduced cytoprotection. PD98059 PD98059 (Figure 6A), suggesting a crucial a essential part of ERK in sulfuretininduced cytoprotection. PD98059 drastically abrogated ERK phosphorylation; however, it did not impact the phosphorylation PD98059 drastically abrogated ERK phosphorylation; having said that, it did not affect the phosphorylation significantly abrogated ERK phosphorylation; on the other hand, it didn’t impact the phosphorylation of Akt of Akt and GSK3 (Figure 6B). collectively, these data suggestsuggest that sulfuretininduced Heneicosanoic acid Epigenetic Reader Domain protective of Akt and (Figure 6B). Taken Taken collectively, these information that sulfuretininduced protective effects and GSK3 GSK3 (Figure 6B). Taken with each other, these information recommend that sulfuretininduced protective effects by AktGSK3ERK pathways are Spermine NONOate supplier independent of each and every each and every other. effects by AktGSK3 and ERK pathways are independent of every other. by AktGSK3 and and ERK pathways are independent of other.Figure 6. Cont.Int. J. Mol. Sci. 2017, 18, 2753 Int. J. Mol. Sci. 2017, 18,11 of 20 11 ofFigure 6. PD98059 suppresses sulfuretininduced protection against MPP SHSY5Y cells have been Figure 6. PD98059 suppresses sulfuretininduced protection against MPP . . SHSY5Y cells were pretreated with without having PD98059 (ten (ten ) for 2 h, followed by remedy with sulfuretin pretreated with or or with out PD98059 ) for 2 h, followed by remedy with or withoutor without having sulfuretin 2 h, and exposed to exposed to MPP (1 h. (A) Cell h. (A) Cell viability was measured by (40 ) for(40 ) for two h, and MPP (1 mM) for 24 mM) for 24viability was measured by MTT assay. MTT are presented are presented relative to manage as mean percentage = three). (B) S.D. (n = three). of Valuesassay. Values relative to manage as imply percentage alter S.D. (nchange Protein levels (B) Protein levels of pERK, ERK, pAkt, Akt, pGSK3, were determined by had been determined by pERK, ERK, pAkt, Akt, pGSK3, GSK3, and GAPDHGSK3, and GAPDHWestern blot analysis. Western blot analysis. Representative blots quantification are shown. Values are are shown. Values Representative blots and their densitometricand their densitometric quantification presented relative are presented relative modify S.D. (n = 3). modify are (n = 3). Differences are statistically to handle as mean fold to manage as mean fold DifferencesS.D. statistically important at p 0.05, important at p 0.05, vs. the control 0.001 p 0.001 vs. the MPP group, the p group, p 0.01, p 0.001 p 0.01, p group,vs. the manage group, p 0.001 vs. and MPP 0.01, and p 0.01, the p 0.001 vs. the MPP pretreated group. p 0.001 vs. MPP and sulfuretin and sulfuretin pretreated group.3. Discussion three. Discussion Naturally occurring flavonoids are polyphenols, located ubiquitously in plants [32]. Naturally occurring flavonoids are polyphenols, found ubiquitously in plants [32]. Interestingly, Interestingly, several flavonoid compounds exhibit neuroprotective effects in In vitro [33]. In vitro quite a few flavonoid compounds exhibit neuroprotective effects in humans [33]. humans experiments experiments have reported that sulfuretin, as a provides neuroprotection against against and have reported that sulfuretin, as a flavonoid, flavonoid, delivers neuroprotection amyloidamyloid and 6OHDA In addition, sulfuretin attenuates neuroinflammation in microglial cells cells [34]. 6OHDA [25,26].[25,26]. Furthermore, sulfuretin attenuates neuroinflammation in microglial[34]. Like L.