![]() There is a non-specific high-molecular-weight band in all lanes in the Na v1.1 blot image. Na v1.1 was not detected in untransduced NgN2 neurons. 12.5 µg of total membrane protein was loaded per sample lane, and the ubiquitous membrane protein Na/K ATPase was used as a sample loading control. ( B) Comparing total expression between Na v1.1-F383S and Na v1.1-F383S-S1328P across two Dox concentrations by western blotting (WB). The mCherry reporter labels transduced neurons, and mCherry signal intensity correlated with Doxycline concentrations. The F383S variant makes the channels resistant to TTX. ( B) hESCs (H9) were first differentiated into a homogeneous neuronal population via NgN2-induction and then transduced with lentiviral vectors to express mCherry-T2A-Na v1.1-F383S or mCherry-T2A-Na v1.1-F383S-S1328P under the control of a Doxycline (Dox) inducible promoter. The corresponding DNA variant (c.3982T>C) was confirmed in patient-derived fibroblasts and induced pluripotent stem cells (iPSCs) by DNA sequencing. ![]() ( A) The p.S1382P mutation is located in Domain III Transmembrane Segment 4 (DIII-S4) of the human Na v1.1 protein. ![]()
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