Rotection against MPP whereas SB415286 Figure 5. LY294002 suppresses sulfuretininduced protection against whereas SB415286 reverses reverses MPPinducedcytotoxicity. SHSYwerecells had been pretreated with or without the need of LY294002 (10 reverses MPPinduced cytotoxicity. cells 5Y pretreated with or with out or without the need of LY294002 (10 MPP induced cytotoxicity. SHSY5YSHSY5Y cells have been pretreated with LY294002 (10 ) for two h, ) for 22h, followed by remedy with or with no (40 ) for(40h andfor22h andto MPP (1 mM)(1 ) for h, therapy with or without having or devoid of sulfuretin two ) exposed exposed to MPP (1 followed by followed by treatment with sulfuretin sulfuretin (40 ) for h andexposed to MPP for mM) for 24 h.viability was measured by MTT assay. MTT assaypresentedare presented relative to 24 h. (A) Cellh. (A) Cell viability was measured by Values are .Values relative to handle as mean mM) for 24 (A) Cell viability was measured by MTT assay .Values are presented relative to control as mean percentage(nchange S.D. (n ==3). (B) Protein levels of pAkt, Akt, pGSK3, GSK3, percentagemean percentagechange (B)S.D. (n levels of pAkt, Akt, of pAkt, Akt, pGSK3, GSK3, Protein 3). (B) Protein levels pGSK3, GSK3, pERK, ERK, handle as adjust S.D. = 3). pERK, ERK, and GAPDH wereby Pyridaben Inhibitor Western blot Conglobatin Purity & Documentation analysis. blot analysis. Representative blots and their and GAPDH wereGAPDH weredetermined by Western Representative blots and theirblots and their pERK, ERK, and determined determined by Western blot evaluation. Representative densitometric quantification quantification are shown. Values relative to manage as to handle as mean fold transform densitometric quantification are shown. Values are presented relativemean fold change old alter densitometric are shown. Values are presented are presented relative to manage as mean S.D. (n = three). (C) SHSY5Y (C) SHSY5Y cells have been pretreated withor without the need of SB415286 (20 ) for two h,then S.D. (n ==3). (C) SHSY5Y cells had been pretreated with SB415286 (20 ) for(20h, andfor2 h,exposed to S.D. (n three). cells had been pretreated with or with out or devoid of SB415286 2 ) then then MPP (1 to MPP (1 mM) for24 h. Cell viability was measuredby MTT assay .Values are presented exposed to MPP(1 mM) for viability was measuredmeasuredassay. Values areValues are presented exposed mM) for 24 h. Cell 24 h. Cell viability was by MTT by MTT assay . presented relative to handle to handle as imply alter S.D. (n = three).S.D. (n 3). Differences are statistically at p 0.01, relative as imply percentage percentage adjust S.D. (n ==3). Differences are statistically important at relative to handle as mean percentage alter Variations are statistically important considerable at p0.01, pp0.001 vs. the manage group, pp0.01, 0.001 0.001 vs. the MPPgroup, p pp 0.001 vs. 0.001 vs. the handle p 0.01, p pp 0.01, the manage group, group, 0.01, ppvs. the vs. the group,group,and 0.01, 0.001 MPP MPP and and and sulfuretin cotreated group. 0.0010.001vs. the MPP and sulfuretin cotreated group. vs. vs. the 0.01,p pp0.001the MPP MPP and sulfuretin cotreated group. 0.01, Moreover, the protective impact of sulfuretin was was abolished within the presenceMAPK inhibitor, Also, the protective impact of sulfuretin was abolished presence with the from the MAPK Also, the protective impact of sulfuretin abolished inside the in the presence of the MAPK inhibitor, PD98059 (Figure 6A), suggesting function of ERK inof ERK in sulfuretininduced cytoprotection. in.