M mlv rt kit

According to manufacturers’ protocol, total RNA was extracted from cells using RNAiso Plus (Takara, 9109). Then, 1 μg of total RNA was reverse transcribed using Evo M-MLV RT kit with gDNA clean for qPCR II (Accurate biology, AG11711) to synthesis cDNA. Relative RT-qPCR was performed using the TransStart^® Top Green qPCR SuperMix (+Dye II) (Transgen, AQ132-24), and target gene expression was calculated by normalization to GAPDH using the 2^–ΔΔCt method. The copy number of the polymerase 3D gene was detected by absolute RT-qPCR using the Premix Ex Taq Probe qPCR (Takara, RR390A). The probe and primer sequences used in this study are described in Table 2.

The total RNA obtained from the purified peripheral γδT cells was converted to complementary DNA (cDNA) using an Evo M‐MLV RT kit with gDNA cleaned for qPCR II (AG111728, Accurate Biotechnology). PCR experiments were performed in a PCR thermal cycler (Bio‐Rad). GAPDH served as the internal control. Primer sequences are listed in Supporting Information: Table 2.