Novel Synthetic Coumarin-Chalcone Derivative (E)-3-(3-(4-(Dimethylamino)Phenyl)Acryloyl)-4-Hydroxy-2H-Chromen-2-One Activates CREB-Mediated Neuroprotection in Aβ and Tau Cell Models of Alzheimer’s Disease

Figure 6

LM-021-mediated kinase activation in Aβ-GFP-expressing SH-SY5Y cells. (a) Experimental flow chart. On day 1, cells were plated with retinoic acid (RA, 10 μM) being added to the culture medium. On day 2, LM-021 (5 μM) was added to the cells for 8 h, followed by treatment of doxycycline (Dox, 5 μg/ml) to induce Aβ-GFP expression. Kinase inhibitors (10 μM) were added to the cells on day 6. On day 8, p-PKA, p-CaMKII, p-ERK, p-CREB, BDNF, BCL, and BAX levels, as well as neurite outgrowth (TUBB3 staining), were measured. (b) p-PKA, p-CaMKII, and p-ERK protein levels determined by immunoblot using GAPDH as a loading control (). To normalize, the protein expression level in untreated cells was set as 100%. (c) Measurements of neurite outgrowth (length, process, and branch) (). values: induced vs. uninduced cells (^#, ^##, and ^###), LM-021-treated vs. untreated cells (, , and ), or kinase inhibitor-treated vs. untreated cells (^&, ^&&, and ^&&&) (one-way ANOVA with a post hoc Tukey test).

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