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. 2018 Feb 27;11:59. doi: 10.3389/fnmol.2018.00059

Figure 4.

Figure 4

Genistein induces the phosphorylation of cAMP response element-binding protein (CREB). (A) Cultured PC12 cells, serum starved for 5 h, were treated with NGF at 20 ng/mL and genistein at 10 μM for different time. Total CREB and phosphorylated CREB (~42 kDa) were revealed by using specific antibodies. (B) Cultured PC12 cells, serum starved for over 5 h, were pre-treated with or without PKA inhibitor, H89 (5 μM) for 3 h prior to the treatment with NGF or genistein for 10 min as in (A). Total CREB and phosphorylated CREB (~42 kDa) were revealed by using specific antibodies (upper panel). Quantification plot was shown in histograms (lower panel). (C) Cultured PC12 cells were transiently transfected with two promoter constructs, pCRE-Luc and pERE-Luc, for 24 h and pre-treated with or without H89. Then, genistein was treated for 2 days. Cell lysates were collected for luciferase assay (left panel). Cultured PC12 cells were transiently transfected with the constructs for 24 h and pre-treated with or without H89 (right panel). Cell lysates were collected for luciferase assay. Data are expressed as the fold of basal value where control value is set as 1, mean ± SEM, n = 5, each with triplicate samples. *p < 0.05; **p < 0.01; ***p < 0.001.