2.3. Gene Expression by qPCR

Gene expression was quantified by qPCR with PerfeCTa SYBR Green master mix as previously described [14]. SIGLEC14 was quantified with primers corresponding to a sequence in exons 3 and 5: 5′-CAGGTGAAACGCCAAGGAG-3′ and 5′-GCGAGGAACAGGGACTGG-3′. SIGLEC5 was quantified with primers corresponding to sequences in exons 4 and 5: 5′-ACCATCTTCAGGAACGGCAT-3′ and 5′-GGGAGCATCACAGAGCAGC-3′. Cycling conditions for all qPCRs were as follows: 95 °C, 2 min; 95 °C, 15 s, 60 °C, 15 s, 72 °C, 30 s, 40 cycles. Copy numbers present in the cDNA were determined relative to standard curves that were executed in parallel [19].