Total RNA extracted from cultured HCAECs or lung endothelial cells (ECs) were performed as previously described20. TA was purchased from Sigma-Aldrich Co. (#MKBV0516V). Cells were grown in 24-well plate and treated with TA (0, 0.1, 1, 10 and 20 µM) for 24 h. Total RNA from cells was extracted using a QIAGEN RNeasy Mini kit (Qiagen) and converted into complementary DNA (cDNA) using a High-Capacity cDNA Reverse Transcription Kit (#4374966, Applied Biosystems, Foster City, CA). Quantitative real-time PCR (qPCR) was performed in a Bio-Rad iQ5 real-time PCR thermal cycler using iQ SYBR Green Supermix (#1708886, Bio-Rad). Relative mRNA expression of target genes was normalized to GAPDH22.