Figure 2: Increased Homer1a protein levels attenuated calcium overload and protected against mitochondrial damage after glutamate treatment.
For long-term [Ca2+]cyt monitoring, after infection with lentivirus particles, the HT-22 cells were transfected with genetically encoded Ca2 + indicator construct (pCMV-B-GECO1) for 24 h. Then, the HT-22 cells were treated with glutamate, and the [Ca2+]cyt was measured at intervals of 1 h for 12 h. The data are represented as time-series curve graph (A). The fluorescence changes were calculated as: total fluorescence value/total cell number. Cells in which the fluorescence signal quenched were excluded from the analysis. To visualize mitochondrial oxidative stress level, HT-22 cells were infected with lentiviral particles for 48 h, followed by transfection with pMitotimer. Twenty-four h after transfection, the HT-22 cells were treated with glutamate for 12 h, and imaged with confocal microscopy. Representative images of mitochondrial oxidative stress are shown (B, Green, non-oxidative levels; Red, oxidative levels), and mitochondrial oxidative levels were quantitated (C). Scale bar = 10 μm. Approximately 30–40 HT-22 cells were analyzed per group. HT-22 cells in the control group were not subjected to lentiviral infection or glutamate injury. Data are presented as mean ± SEM from four experiments. *p< 0.05 vs. control group; #p < 0.05 vs. control + glutamate or vector + glutamate group.
