A Role of Canonical Transient Receptor Potential 5 Channel in Neuronal Differentiation from A2B5 Neural Progenitor Cells
Figure 5
Effect of 2-APB on TG-stimulated Ca2+-influx and cell viability in differentiated neuronal cells.
The A2B5+ NPCs were cultured in NM media supplemented with 10, 50 and 100 µM 2-APB for five days, and then the cells were switched to media without 2-APB. (A): After 3 weeks, TG-stimulated Ba2+ entry was monitored in A2B5+ NPCs and differentiated neuronal cells. After store depletion, TG-stimulated Ba2+ influx was inhibited by the addition of 2-APB in differentiated media. Each trace is the average of A2B5+ NPCs or differentiated neuronal cells. The time of Ba2+ addition is indicated by the dark grey and that of 2-APB is done by the grey. Error bars represent SE. The data are representative of at least 3 separate experiments. (B): After 3 weeks, the A2B5+ NPCs treated with 10 µM 2-APB normally differentiated into mature neuronal cells. However, the rate of differentiation of the cells significantly reduced in the media supplemented with 50 µM 2-APB, and the cells hardly differentiated into mature neuronal cells in the media supplemented with 100 µM 2-APB. (C): The proliferation assay was done to show effects of SOCE inhibitor on the fate determination and survival of A2B5+ NPCs in the differentiation condition. For proliferation assay, the A2B5+ NPCs were treated with 2-APB at the indicated concentrations (10, 50 and 100 µM) 24 h after plating. Data was expressed as the mean optical density of 3 readings. Error bars indicate the standard deviation. Scale bars, 50 µm.