2.7. Real-Time Quantitative PCR
SUDHL4 and Farage cells were treated with compounds for 24 h and lysed
with TRIzol reagent (Invitrogen). RNA was extracted and reverse
transcribed into cDNA with the Prime Script RT Reagent Kit (Takara). The
cDNA was then used as the template for the RT-qPCR reaction and the
RT-qPCR reaction was performed using SYBR-Green (Takara) on
QuantStudio®3 Real-Time PCR System (Applied Biosystems). The sequences
of primers used in the study were as follows:
p53: CCCTTCCCAGAAAACCTACC and AATCAACCCACAGCTGCAC,
CDKN1A: CTGAAGGGTCCCCAGGTG and TAGGGCTTCCTCTTGGAGAA,
CXCR4: AGGCCCTAGCTTTCTTCCAC and CTGCTCACAGAGGTGAGTGC,
CD69: CTGGTCACCCATGGAAGTG and CATGCTGCTGACCTCTGTGT,
β-actin: TGAAGTGTGACGTGGACATC and CATACTCCTGCTTGCTGATC.