Name | Sequence |
---|---|
NKCC1a (1-27) forward primer | 5′-ACCAAGGATGTGGTAGTAAGTGTGG-3′ |
NKCC1a (1-27) reverse primer | 5′-GGATTCTTTTTTCAACAGTGGTTGA-3′ |
NKCC1a (1-27) probe | 6FAMTACACACAAAGTTGAGGAAGMGBNFQ |
NKCC1b (1-27 (Δ21)) forward primer | 5′-AGGATGTGGTAGTAAGTGTGGAATATAGTAA-3′ |
NKCC1b (1-27 (Δ21)) reverse primer | 5′-TCAAGAAGCTTTTGGTCAGCTACAT-3′ |
NKCC1b (1-27 (Δ21)) probe | 6FAMTACACACAAAGAATCCAMGBNFQ |
NKCC1 1-27 (21a) forward primer | 5′-GAATATAGTAAAAAGTCCGATTTAGATACTTCC-3′ |
NKCC1 1-27 (21a) reverse primer | 5′-CCTTCTCATAAAATTCATGTATGTTGACA-3′ |
NKCC1 1-27 (21a) probe | 6FAMTTACACACAAAGTCGTGCTGMGBNFQ |
NKCC1 1-4a forward primer | 5′-TGACAACTATCACAGGATTGTCTACTTC-3′ |
NKCC1 1-4a reverse primer | 5′-ATTCTTTTCTTCTTGTTTTCAGGGATC-3′ |
NKCC1 1-4a probe | 6FAMAGAGGAGGAAGAGGACMGBNFQ |
NKCC1 1-2a forward primer | 5′-CTGTGGTCACGTATACTGCAGAAAGTA-3′ |
NKCC1 1-2a reverse primer | 5′-AACTGATGAGAACGTTAGGAAGATTTTATA-3′ |
NKCC1 1-2a probe | 6FAMCAAGGGTGTATTAGTATGTATATMGBNFQ |
↵aWe assessed mRNA expression levels of novel NKCC1 transcripts [isoforms 1–27 (21a), 1–4a, and 1–2a] as well as the known transcripts NKCC1a (1–27) and NKCC1b (1–27 (Δ21)) in the DLPFC by qRT-PCR (Table 3). We measured the expression of NKCC1a (1–27), using an assay spanning exon 20 and exon 21, expression of NKCC1b (1–27 (Δ21)), using an assay spanning exon 20 and exon 22, expression of 1–4a, using an assay spanning exon 4 and a unique exon 4a, and expression of NKCC1b 1–2a, using an assay spanning exon 2 and a unique exon 2a. We did not measure the isoform 1–27 (Δ5–26) by qPCR because of low abundance (Ct > 36) of mRNA expression in our preliminary study (data not shown). All data were normalized to a geometric mean of three “housekeeping genes.”