2.10 | Quantitative RT-PCR (qRT-PCR)
We performed qRT-PCR to determine the expression of MG53, TGF-β1,
Smad2/3, ZO-1, and Col-a1 in treated and untreated nasal tissues and
epithelial cells. TRIZOL reagent (TAKARA, Japan) was used to extract
total RNA; contamination by genomic DNA was removed using gDNA Eraser
(TAKARA, Japan), following the manufacturer’s instructions. Next, total
RNA (1000 ng) was reverse transcribed by cDNA synthesis (TAKARA,
Japan). qRT-PCR was performed using a
SYBR Premix ExTaq II kit (TAKARA, Japan) with a CFX96 RT-PCR Detection
System (Bio-Rad, Hercules, USA). Human GAPDH gene was used as the
internal control. The primers used in this study are shown in Table 1.