Proliferating K562 RepID WT and KO cells were harvested, and RNA was extracted using the RNeasy R Mini Kit (Qiagen, 74104) and converted to cDNA by using PrimeScript RT Master Mix (Takara, RR036A) according to the manufacturer’s instructions. An RT2 Profiler PCR array (Qiagen, PAMM-043Z) with 84 genes associated with the Wnt signaling pathway was used.
Total RNA extraction, cDNA synthesis, and qRT-PCR were performed according to the manufacturer’s instructions provided by Qiagen (RNeasy R Mini Kit, 74104), Takara (PrimeScript RT Master Mix RR036A), and Invitrogen (SYBR Green PCR Master Mix 4309155), respectively. The expression levels of human DAB2, CD41, and CD61 in the cells were determined by qRT-PCR, using GAPDH as an internal control. DAB2 primers; forward:5’-CCA GAT GCA AGA GGG GAT AA-3’ and reverse:5’-TCC TCC ACA CAC GTA ACC AA-3,’ CD41 primers; forward:5’-AGG TGA GAG GGA GCA GAA CA-3’ and reverse:5’-TCC ACC TTG AGA GGG TTG AC-3,’ CD61 primers; forward:5’-GAC AAG GGC TCT GGA GAC AG-3’ and reverse:5’-ACT GGT GAG CTT TCG CAT CT-3,’ GAPDH primers; forward:5’-GAG TCA ACG GAT TTG GTC GT-3’ and reverse:5’-TTG ATT TTG GAG GGA TCT CG-3.’
Total RNA extraction, cDNA synthesis, and qRT-PCR were performed according to the manufacturer’s instructions provided by Qiagen (RNeasy R Mini Kit, 74104), Takara (PrimeScript RT Master Mix RR036A), and Invitrogen (SYBR Green PCR Master Mix 4309155), respectively. The expression levels of human DAB2, CD41, and CD61 in the cells were determined by qRT-PCR, using GAPDH as an internal control. DAB2 primers; forward:5’-CCA GAT GCA AGA GGG GAT AA-3’ and reverse:5’-TCC TCC ACA CAC GTA ACC AA-3,’ CD41 primers; forward:5’-AGG TGA GAG GGA GCA GAA CA-3’ and reverse:5’-TCC ACC TTG AGA GGG TTG AC-3,’ CD61 primers; forward:5’-GAC AAG GGC TCT GGA GAC AG-3’ and reverse:5’-ACT GGT GAG CTT TCG CAT CT-3,’ GAPDH primers; forward:5’-GAG TCA ACG GAT TTG GTC GT-3’ and reverse:5’-TTG ATT TTG GAG GGA TCT CG-3.’