Quantifying Transcription Factors in Lung Tissue

\The transcription factors of IFN-γ, IL-4, FOXP3, and RORγt in lung tissue were detected, and the total RNA from the tissues was extracted by TRIzol (Invitrogen) in accordance with the manufacturer’s instructions. cDNA was synthesized by reverse transcription by using the first-strand cDNA synthesis kit (Servicebio, G3330) in accordance with the manufacturer’s instructions and used for real-time PCR assay performed in 1× SYBR green qPCR master mix (Servicebio, G3320) together with 0.2 mM forward and reverse primers. The amount of mRNA of the indicated genes after normalization of β-actin mRNA. The primers of GAPDH, Foxp3, RORγt, IFN-γ and IL-4 genes were as described as in a previous study (Zhou et al., 2022 (link)).