DRG were dissected as previously described (refer to DRG extraction section) and were collected in an Eppendorf tube containing 200 μL/tube of RNA-Later solution. Tubes were left at −20 °C overnight and the day after tissue was crushed with a pestle and lysed by adding 200 μL/tube of freshly prepared lysis buffer (RNeasy Protect Mini Kit, Qiagen) and mercaptoethanol. RNA was extracted following standard procedure using the RNeasy Protect Mini protocol (Qiagen) including DNase I digestion step. Total RNA extracted was converted to first-strand cDNA using the SuperScript III First-Strand Synthesis System kit (Thermo Fisher Scientific). For HEK-293 cell experiment, cells were lysed by adding 600 μL/well of freshly prepared lysis buffer (RNeasy Protect Mini Kit, Qiagen) and mercaptoethanol. Cell lysates were transferred to Eppendorf tubes and RNA was extracted following standard procedure using the RNeasy Protect Mini protocol (Qiagen) including DNase I digestion step. Total RNA extracted was converted to first-strand cDNA using the SuperScript® III First-Strand Synthesis System kit (Thermo Fisher Scientific).
De Virgiliis F., Hutson T.H., Palmisano I., Amachree S., Miao J., Zhou L., Todorova R., Thompson R., Danzi M.C., Lemmon V.P., Bixby J.L., Wittig I., Shah A.M, & Di Giovanni S. (2020). Enriched conditioning expands the regenerative ability of sensory neurons after spinal cord injury via neuronal intrinsic redox signaling. Nature Communications, 11, 6425.
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