2.5. Quantitative real-time PCR (qRT-PCR)

Cells were lysed and total RNA was isolated from cell lysates using an RNeasy kit (Qiagen; Valencia, CA, USA) according to the manufacturer’s directions. RNA was reverse transcribed to cDNA using the High-Capacity cDNA Archive Kit (Applied Biosystems; Carlsbad, CA, USA). The target genes (TNFA, IL1β, IL6, IL8 (CXCL8), CCL8, CXCL10, CCL17, ANGPTL4) vs ACTB or 18S were amplified by qRT-PCR using gene-specific TaqMan Gene Expression Assays (Applied Biosystems). Primer ID and target exons are included in Table 1. Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels.

Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.