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 2

CRE fluorescence reporter assay. (a) Experimental flow chart to determine optimal Ca²⁺ ionophore treatment time. CRE-GFP 293 cells were treated with Ca²⁺ ionophore (2 μM) for 0–24 h. Cell viability was assessed by MTT assay (; two-tailed Student’s -test; and ). (b) Left: experimental flow chart to determine optimal Ca²⁺ ionophore treatment concentration. CRE-GFP 293 cells were treated with Ca²⁺ ionophore (0.5–10 μM) for 5 h. GFP fluorescence was assessed by flow cytometry (; two-tailed Student’s -test; ). Right: Flp-In CRE fluorescence reporter cells with or without Ca²⁺ ionophore (2 μM) treatment for 5 h. (c) Fluorescence analysis of CRE reporter cells untreated or treated with Ca²⁺ ionophore (2 μM) and forskolin, LM-016, LM-021, or LM-022 (5–10 μM) for 5 h. GFP fluorescence was assessed by flow cytometry (; one-way ANOVA with a post hoc Tukey test). values: compound treated vs. untreated cells ( and ), or 10 μM compound-treated vs. 5 μM compound-treated cells (^&&&).

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