Table 1.
Primers used to amplify cDNAs based on published GenBank sequences for mice. bp, base pairs; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; ICAM-1, intercellular adhesion molecule-1; iCreER, tamoxifen inducible Cre recombinase, MCP-1, monocyte chemoattractive protein-1; MIP-2, macrophage inflammatory protein-2; Pdgfb, platelet derived growth factor-b; S1PR, sphingosine 1-phosphare receptor; SK, sphingosine kinase; TNF-α, tumor necrosis factor-alpha; HSP, heat shock protein; HO-1, heme oxygenase-1.
| Primers | Species | Sequence (Sense/Antisense) | Product Size (bp) |
Cycle Number |
Annealing Temp (°C) |
|---|---|---|---|---|---|
| S1P1R | Mouse | 5 ′-CGGTGT AGACCC AGAGTCCT-3′ 5′-AGC AGC AGATGAGAATGAAC-3′ |
393 | 20 | 64 |
| S1P2R | Mouse | 5′- AAAACC AACC ACTGGCTGTC-3′ 5 ′-GAGTGGAACTTGCTGTT-3′ |
317 | 25 | 60 |
| S1P3R | Mouse | 5′-AAGCCTAGCGGGAGAGAAAC-3′ 5 ′-GGCAATC AAAACC ATCAGGT-3′ |
452 | 22 | 64 |
| S1P4R | Mouse | 5 ′-GC AGAAGTCTCC ACGTCCTC-3′ 5 ′-GCTGAGTGACCGAGAAGTCC-3′ |
403 | 23 | 62 |
| S1P5R | Mouse | 5′-ACACCAAATGCCCAGCTTAC- 3′ 5′- ACC AAGAGC AC AGCC AAGTTC-3′ |
335 | 32 | 62 |
| S1P1R FLOX | Mouse | 5′-GAGCGGAGGAAGTTAAAAGTG- 3′ 5′-CCTCCTAAGAGATTGCAGCAA- 3′ |
Flox 250, Cont 200 |
45 | 56 |
| PdgfbiCreER | Mouse | 5′-CCAGCCGCCGTCGCAACT- 3′ 5′-GCCGCCGGGATCACTCTCG- 3′ |
450 | 35 | 60 |
| TNF-α | Mouse | 5 ′-TACTGAACTTCGGGGTGATTGGTCC-3′ 5 ′-C AGCCTTGTCCCTTGAAGAGAACC-3′ |
290 | 24 | 65 |
| ICAM-1 | Mouse | 5 ′-TGTTTCCTGCCTCTGAAGC-3′ 5 ′-CTTCGTTTGTGATCCTCCG-3′ |
409 | 21 | 60 |
| MCP-1 | Mouse | 5′- ACCTGCTGCTACTC ATTC AC-3′ 5 ′-TTGAGGTGGTTGTGGAAAAG-3′ |
312 | 22 | 60 |
| MIP-2 | Mouse | 5 ′-CC AAGGGTTGACTTC AAGAAC-3′ 5′-AGCGAGGC AC ATC AGGTACG-3′ |
282 | 28 | 60 |
| HSP27 | Mouse | 5 ′-CCTAAGGTCTGGC ATGGT A-3′ 5′-AGGAAGCTCGTTGTTGAAGC-3′ |
373 | 25 | 66 |
| HSP27 | Human | 5 ′-GTCCCTGGATGTC AACC AC-3′ 5 ′-GACTGGGATGGTGATCTCG-3′ |
286 | 15 | 65 |
| HSP32 (HO-1) |
Mouse and human | 5 ′-TGAAGGAGGCC ACC AAGGAG-3′ 5 ′-GTGGGCC ACC AGC AGCTC-3′ |
320 | 15 | 65 |
| HSP70 | Mouse and human | 5 ′-GATC ACC ATC ACC AACGAC AA-3′ 5 ′-TTGTCC AGC ACCTTCTTCTTGTC-3′ |
218 | 15/18 | 65(H)/ 60(M) |
| GAPDH | Mouse and human | 5’-ACCACAGTCCATGCCATCAC-3’ 5’-CACCACCCTGTTGCTGTAGCC-3’ |
450 | 15 | 65 |
Respective anticipated PCR product size (bp, base pairs), PCR cycle number for linear amplification, and annealing temperatures used for each primer are provided.