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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Glia. 2014 Apr 17;62(7):1162–1175. doi: 10.1002/glia.22671

Figure 7.

Figure 7

Nav1.5 mRNA knockdown attenuates astrocytic [Ca2+]i response after injury. (a) After scratch, there was a robust [Ca2+]i response in cells treated with NT siRNA, which was propagated through the syncytium of astrocytes and slowly resolved after 2-4 min (first column; see also (b) and Video 4). Knockdown with Nav1.5 siRNA attenuated the [Ca2+]i response (second column, see also Video 5) compared to NT siRNA. Color scale represents the ratio of fluorescent signals induced by 340 and 380 nm excitation in cells loaded with Fura-2 AM. Scale bar 50 µm. (b) Nav1.5 siRNA attenuated the initial [Ca2+]i increase after injury, particularly within the first 60 s. (c) Compared to NT siRNA control (n=5 experiments, 50 cells total), Nav1.5 siRNA treatment (n=3 experiments, 30 cells total) decreased the 60 s AUC by 18 ± 2%, and (d) decreased the peak [Ca2+]i by 18 ± 3%. (e) Quantitative real-time RT-PCR confirmed that Nav1.5 mRNA expression of cells treated with Nav1.5 siRNA was decreased by 68 ± 5% compared to NT siRNA, indicating successful knockdown of Nav1.5. All data was taken from cells within the first 50 µm (3-4 cells deep) from the scratch. *p<0.05.