Figure 2
Effects of LDH-A inhibition on U87MG cell apoptosis, cell cycle progression and Δψm.
(a) U87MG cells were treated for 48 h with DMSO (control), or NHI-1 or NHI-2 at 1 μM or 10 μM. At the end of the treatment periods, the cells were collected and the level of phosphatidylserine externalisation was evaluated using the Annexin V-staining protocol. The data were expressed as the percentage of apoptotic cells versus the total number of cells and are the mean ± SEM of three different experiments. (b) U87MG cells were treated for 72 h with DMSO (control), or NHI-1 or NHI-2 at 1 μM or 10 μM, or DCA at 100 μM and the cell cycle was analysed. The data were expressed as percentage of cell in the different phases (G0/G1, G2/M or S) versus total cell number and they are the mean values ± SEM of three different experiments. (c,d) U87MG were treated for 48 h with DMSO (control) or 50 μM NHI-1 or NHI-2. At the end of treatments, the mitochondria were isolated and the Δψm (5 μg of proteins) was evaluated using JC-1 protocol (c) Representative graph of the mitochondrial potential analysis using the JC-1 protocol. (d) The data were expressed as the variation of JC-1 uptake into mitochondria, which was calculated as the difference between the RFU at the beginning and those read after 10 min and represent the means ± SEM of three different experiments, each performed in duplicate. The significance of the differences was determined by one-way ANOVA and Bonferroni’s post hoc test: *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 vs. the control.
