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. 2016 Oct 17;14(10):187. doi: 10.3390/md14100187

Figure 3.

Figure 3

Effect of 11-de on p-CREB, and BDNF expression in 6-OHDA-treated SH-SY5Y cells. SH-SY5Y were pretreated with 10 nM 11-de for 1 h and 20 μM 6-OHDA plus 11-de for 3 h. Western blotting of p-CREB (A) in control, 6-OHDA-treated, 6-OHDA plus 11-de-treated, and 11-de-treated groups (n = 3). The expression of p-CREB was significantly upregulated after the pretreatment with 10 nM 11-de for 1h and 6-OHDA plus 11-de for 3 h in cells. Detection of the effect of 11-de on 6-OHDA-induced downregulation of CRE binding activity in SH-SY5Y cells in electrophoretic mobility shift assay (EMSA) analysis (B). The upper red rectangle indicates p-CREB, and the lower arrow indicates the free probe. EMSA of control, 6-OHDA-treated, and 6-OHDA plus 11-de-treated groups, respectively (n = 3). Treatment with 11-de significantly increased p-CREB expression in the SH-SY5Y cells pretreated with 10 nM 11-de for 1 h, and cells pretreated with 6-OHDA plus 11-de for 3 h. We further examined the effect of 11-de on brain-derived neurotrophic factor (BDNF) expression after 6-OHDA administration. SH-SY5Y cells were pretreated with 10 nM 11-de for 1 h and incubated with 6-OHDA for 15 h. (C) Immunofluorescence staining of BDNF (red) in the control, 20 μM 6-OHDA 15 h treatment, 6-OHDA plus 11-de-treated, and 11-de-treated groups (n = 3). Treatment with 11-de significantly upregulated BDNF expression both in the presence and in the absence of 6-OHDA administration.