Quantifying vTR and hTR Expression in MDV-Infected Cells

vTR and hTR expression levels were quantified in MDV-infected cells and tumor tissues using RT-qPCR. Briefly, 10⁶ CECs were infected with 10³ PFU of wt, v∆vTR, or vhTR. Total RNA was isolated from infected CECs and tumor tissues using the RNeasy Plus Mini Kit (Qiagen) and the innuPREP Virus RNA kit (Analytik Jena GmbH), respectively, according to the manufacturer’s instructions. The isolated RNAs were treated with RQ1 RNase-Free DNase (Promega), and reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). vTR and hTR expression was quantified using TaqMan qPCR, and the cellular GAPDH reference gene was used to normalize vTR and hTR expression. The vTR, hTR, and GADPH primer and probe sets have been previously published (14 (link), 21 (link)) and are shown in Table 1.