Abi prism 7900pcr instrument

TRIzol reagent and miRNA reverse transcriptase kit were purchased from Invitrogen; Thermo Fisher Scientific, Inc. (Waltham, MA, USA); SYBR Green Master Mix (Applied Biosystems; Thermo Fisher Scientific, Inc.) and ABI Prism 7900PCR instrument was purchased from Applied Biosystems; Thermo Fisher Scientific, Inc., and the reverse transcription primers and internal reference primers were synthesized by Shanghai Biological Engineering Co. Ltd. (Shanghai, China).

TRIzol, 2X SYBR-Green qPCR mix, RevertAid™ Premium First Strand cDNA Synthesis kit, Hank's liquid, penicillin-streptomycin double antibodies and PBS were purchased from Invitrogen; Thermo Fisher Scientific, Inc. (cat. nos. 15596018, 4309155, K1621, 88284, 15070063 and 10010049; Waltham, MA, USA); reverse transcription kit from Takara Biotechnology Co., Ltd. (RNA to cDNA EcoDry™ Premix Double Primed; cat. no. 639549; Beijing, China); quantitative PCR kit from Takara Bio, Inc. (Otsu, Japan); CD69 monoclonal antibody (mouse anti-human phycoerythrin marker), CD3 monoclonal antibody (mouse anti-human activating protein), CD4 monoclonal antibody (BD Horizon™ BV510 mouse anti-human) and PE-labeled mouse IgG1 CD69 isotype control from BD Biosciences (dilution, 1:400; cat. nos. 560738, 552852, 562971 and 557908; Franklin Lakes, NJ, USA); erythrocyte splitting liquor from Tiangen Biotech Co., Ltd. (cat. no. RT122; Beijing, China); ABI Prism 7900 PCR instrument from Applied Biosystems (Thermo Fisher Scientific, Inc., Foster City, CA, USA), BD FACSCanto™ II flow cytometry from BD Biosciences.