Figure 2.

Erinacine A prevention of 1-methyl-4-phenylpyridinium (MPP⁺)-induced cell death and reactive oxygen species (ROS) generation of Neuro-2a (N2a) cells. (A) Cell apoptosis of the N2a cell line treated with 0.1% dimethyl sulfoxide (DMSO), MPP⁺, or MPP⁺ and erinacine A (1 and 5 µM) at 24 h measured by DAPI staining under fluorescence microscopy. The red arrows indicate the condensed apoptotic cells with fragmented nuclei at a magnification of ×200. (B) Flow cytometry analysis of the N2a cell line treated with 0.1% DMSO, MPP⁺, or MPP⁺ and erinacine A (1 and 5 µM) at 24 h stained by fluorescein isothiocyanate (FITC)-conjugated annexin V and propidium iodide (PI). The percentages of these treated cells with positive annexin V and/or PI and their changes (fold increase of untreated control) are shown in each quadrant. (C) After being incubated with H2DCFDA, the intracellular ROS of the N2a cell line treated with 0.1% DMSO, MPP⁺, or MPP⁺ and erinacine A at 24 h was detected by FACS analysis. Typical FACS profiles for H2DCFDA in the cells under different treatments are shown in the upper panel; the fluorescence intensity of each treatment and their changes (fold increase of untreated control) are shown in the bottom table.