Green premix ex taqtm 2

All the animals were sacrificed in deep anesthesia with isoflurane. For total RNA extraction of tissue, the lumbar enlargement of the spinal cord was removed and homogenized with Trizol (Life Technologies, United States). The total RNA was subjected to reverse transcription using PrimeScript^TM RT reagent Kit with gDNA Eraser (Takara, Japan), followed by qPCR using Green^TM Premix Ex Taq^TM II (Takara, Japan). For RNA extraction of sorted microglia, microglia were homogenized with Trizol (Life Technologies, United States). Then, the RNA was subjected to dissolution, denaturation, reverse transcription, and amplification as described (Chen et al., 2017 (link)). qPCR was performed using Green^TM Premix Ex Taq^TM II (Takara, Japan). The primers are shown in Table 1. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) functioned as the endogenous control gene. Messenger RNA (mRNA) expressions were analyzed according to the 2^–ΔΔCT method.