Table 1.
Primers used to amplify mRNAs encoding SK1, SK2, GAPDH, and proinflammatory cytokines based on published GenBank sequences for mice.
| Primers | Accession Number | Sequence (Sense/Antisense) | Product Size (bp) | Cycle Number | Annealing Temp (°C) |
|---|---|---|---|---|---|
| Mouse SK1 | NM_011451 (variant 1) | 5'–GATGCATGAGGTGGTGAATG–3' | 337 | 22 | 64 |
| NM_025367 (variant 2) | 5'–GCCCACTGTGAAACGAATC–3' | ||||
| Mouse SK2 | NM_203280 (variant 1) | 5'–ACTGCTCGCTTCTTCTCTGC–3' | 437 | 23 | 68 |
| NM_020011 (variant 2) | 5'–ACCATTGAGGGACAGGTCAG–3' | ||||
| Mouse MIP-2 | X53798 | 5'–CCAAGGGTTGACTTCAAGAAC–3' | 282 | 28 | 60 |
| 5'–AGCGAGGCACATCAGGTACG–3' | |||||
| Mouse ICAM-1 | X52264 | 5'–TGTTTCCTGCCTCTGAAGC–3' | 409 | 21 | 60 |
| 5'–CTTCGTTTGTGATCCTCCG–3' | |||||
| Mouse TNF-α | X02611 | 5'–TACTGAACTTCGGGGTGATTGGTCC–3' | 290 | 24 | 65 |
| 5'–CAGCCTTGTCCCTTGAAGAGAACC–3' | |||||
| Mouse MCP-1 | NM_011333 | 5'–ACCTGCTGCTACTCATTCAC–3' | 312 | 22 | 60 |
| 5'–TTGAGGTGGTTGTGGAAAAG–3' | |||||
| Mouse IL-6 | NM_031168 | 5'–CCGGAGAGGAGACTTCACAG–3' | 421 | 30 | 62 |
| 5'–GGAAATTGGGGTAGGAAGGA–3' | |||||
| Mouse IL-17 | NM_010552 | 5'–TCCAGAAGGCCCTCAGACTA–3' | 248 | 32 | 66 |
| 5'–ACACCCACCAGCATCTTCTC–3' | |||||
| Mouse GAPDH | M32599 | 5'–ACCACAGTCCATGCCATCAC–3' | 450 | 15 | 65 |
| 5'–CACCACCCTGTTGCTGTAGCC–3' |
bp, base pairs; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; ICAM-1, intercellular adhesion molecule-1; MCP-1, monocyte chemoattractant protein 1; MIP-2, macrophage inflammatory protein 2; SK, sphingosine kinase; TNF-α, tumor necrosis factor alpha. Respective anticipated RT-PCR product size, PCR cycle number for linear amplification and annealing temperatures used for each primer are also provided.