CXCR4 Gene Expression Quantification

We employed RNeasy reagent (Qiagen, Germany) to harvest the whole RNA. RT-qPCR was conducted with SYBR Premix ex TAG Mastermix kit (Takara, Japan) on the ICycler real-time system (Bio-Rad Laboratories, USA) as manual described. Glyceraldehyde-3-phosphate dehydrogenase was an internal control. The relative RNA expression was analyzed by the 2^−ΔΔCt approach and presented as the target gene/internal control ratio [2^{−ΔΔCt (target gene-internal control)}] (28 (link)). The data were obtained from three independent experiments in triplicate one time. The primers of CXCR4 are 5’-ACTACACCGAGGAAATGGGCT-3’ (F) and 5’-CCCACAATGCCAGTTAAGAAGA-3’ (R). The primers of GAPDH are 5’-CTGGGCTACACTGAGCACC-3’ (F) and 5’-CTGGGCTACACTGAGCACC-3’ (R).