Quantitative PCR
Total messenger RNA (mRNA) was extracted from lung tissues using TRIzol
regent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s
instructions and then reverse transcribed into
complementary DNA (cDNA) using
the TIANGEN kit (KR 103, TIANGEN Biotechnology, Beijing, China). An SYBR
Green I Q-PCR kit (TransGen Biotech, Beijing, China) was used to amplify
PCR by the Bio-Rad IQ5 system (Bio Rad, Hercules, CA, USA ). A
fluorescence reporter signal was detected based on the internal
reference dye signal for β -actin to normalize for non-PCR
associated fluorescence fluctuations between microwells. Supplementary
Table 1 lists the primer sequences used in our experiments. All primers
were synthesized by Beijing Tianyihuiyuan Biotechnology Company.