Platinum sybr green qpcr mix

PP total RNA was purified using the RNeasy Mini Kit (Qiagen) and cDNA was synthesized using the M-MLV Reverse Transcriptase (Promega). Real-time (RT) PCR was performed in the CFX96 Real Time System (Bio-Rad) according to the manufacturer’s instructions, using the Platinum SYBR Green qPCR Mix (Thermo Fisher) and with the following primers: Gp2-F 5′-CTGCTACCTCGAAGGGGACT-3′; Gp2-R 5′-CATTGCCAGAGGGAAGAACT-3′; Spib-F GCCCACACTTAAGCTGTTTGTA-3′; Spib-R 5′-CTGTCCAGCCCCATGTAGAG-3′; Ccl20-F 5′-GATGGCCGATGAAGCTTGTG-3′; Ccl20-R 5′-GACTCTTAGGCTGAGGAGGTTC-3′; Rankl-F 5′-AAACAAGCCTTTCAGGGGGC-3′; Rankl-R 5′-TCCAACCATGAGCCTTCCATC-3′; Ccl19-F 5′-CCTTAGTGTGGTGAACACAACA-3′, Ccl19-R 5′-GGGTGCTAATGATGCGGAA-3′, Ccl21-F 5′-GCTGCAAGAGAACTGAACAGACA-3′, Ccl21-R 5′-CGTGAACCACCCAGCTTG-3′, Cxcl13-F 5′-TCTCTCCAGGCCACGGTATTCT-3′ and Cxcl13-R 5′-ACCATTTGGCACGAGGATTCAC-3′, b2m-F 5′-TTCTGGTGCTTGTCTCACTGA-3′, b2m-R 5′-CAGTATGTTCGGCTTCCCATTC-3′. Data were normalized to the housekeeping gene (b2m), analyzed using the 2^–ΔΔCT method, and expressed relative to control groups as indicated.