Faraguna_Pena_Kantarci_STAR Protocols

RNAscope images shows the expression of Nav1.6 (C3), Nav1.7(C1) and Nav1.8(C2) in DRG neurons isolated using the protocol described here, cultured for 9 days in vitro (DIV), and either treated with DMSO (0.01%) or 1 uM 16,16-dimethyl PGE2 (dmPGE2)—a PGE2 derivative that is resistant to degradation on DIV8 for 16-24 h. dmPGE2 increases the expression of indicated voltage-gated sodium channels as discussed in our referenced publication ( PMID: 39142281). These images are added in relation to our protocol that describes purification of dorsal root ganglion neurons in highly pure cultures. Isolated DRG neurons can be stained using in situ hybridization, RNAscope, as shown in these example images.

RNAscope图像展示了Nav1.6（C3）、Nav1.7（C1）及Nav1.8（C2）在背根神经节（Dorsal Root Ganglion，DRG）神经元中的表达模式。这些神经元经本文所述方案分离后，体外培养9天（days in vitro，DIV），并于DIV8时分别用0.01%二甲基亚砜（Dimethyl Sulfoxide，DMSO）或1 μM 16,16-二甲基前列腺素E2（16,16-dimethyl Prostaglandin E2，dmPGE2）处理16-24小时——dmPGE2为一种抗降解的前列腺素E2（Prostaglandin E2，PGE2）衍生物。正如我们引用的文献（PMID: 39142281）所讨论，dmPGE2可上调上述电压门控钠通道的表达水平。这些图像补充了我们关于高纯度培养体系中DRG神经元纯化方案的描述。示例图像表明，分离的DRG神经元可通过原位杂交技术（RNAscope）进行染色。