Screening for genes expressed in E11.5 3PPs
| Gene from subtraction screening . | Verification of expression specificity . | ||||
|---|---|---|---|---|---|
| Group/No. . | Name . | Accession no. . | Frequency . | 3PP/2PA (SE) . | . |
| 1 | |||||
| 1 | CCL21 | NM_011124 | 254 | 407.7 (86) | * |
| 2 | PTH | NM_020623 | 104 | 2.1 (1.1) | * |
| 3 | Casr | NM_013803 | 10 | 19 088.5 (1963) | * |
| 4 | Spink8 | NM_183136 | 8 | 53.9 (19) | * |
| 5 | Pla2g7 | NM_013737 | 7 | 5.3 (3.8) | * |
| 6 | Elovl2 | NM_019423 | 6 | 0.5 (0.3) | |
| 7 | MafB | NM_010658 | 6 | 1 842.7 (542) | * |
| 8 | Irs4 | NM_010572 | 5 | 4 166.0 (958) | * |
| 9 | IL7 | NM_008371 | 4 | 658.7 (32) | * |
| 10 | Nedd4 | NM_010890 | 3 | 210.3 (94) | * |
| 11 | Pax1 | NM_008780 | 3 | 2.5 (2.1) | * |
| 12 | Gcm2 | NM_008104 | 3 | 8.2 (4.6) | * |
| 13 | Angpt2 | NM_007426 | 3 | 3.2 (0.7) | * |
| 14 | Eeflg | NM_026007 | 3 | 0.1 (0.1) | |
| 15 | 2610528KiiRik | NM_175184 | 3 | 0.8 (0.1) | |
| 16 | Gpsn2 | NM_134118 | 3 | 0.3 (0.1) | |
| 17 | Hnrnpf | NM_133834 | 3 | 0.5 (0.1) | |
| 17 genes | 12 genes | ||||
| 2 | |||||
| 1 | Dtprp | NM_010088 | 2 | 0.1 (0.0) | |
| 2 | Pbx1 | NM_008783 | 2 | 1.6 (0.7) | |
| 3 | Atp5a1 | NM_007505 | 2 | 0.4 (0.1) | |
| 4 | Zfp9 | NM_011763 | 2 | 1.1 (0.5) | |
| 50 genes | |||||
| 3 | |||||
| 1 | Ndufv1 | NM_133666 | 1 | 0.1 (0.0) | |
| 2 | Neurod | NM_010894 | 1 | 0.3 (0.1) | |
| 3 | Cks2 | NM_025415 | 1 | 0.3 (0.1) | |
| 4 | Ptpn18 | NM_011206 | 1 | 0.1 (0.0) | |
| 150 genes | |||||
| Total | 217 genes | 12 genes | |||
| Gene from subtraction screening . | Verification of expression specificity . | ||||
|---|---|---|---|---|---|
| Group/No. . | Name . | Accession no. . | Frequency . | 3PP/2PA (SE) . | . |
| 1 | |||||
| 1 | CCL21 | NM_011124 | 254 | 407.7 (86) | * |
| 2 | PTH | NM_020623 | 104 | 2.1 (1.1) | * |
| 3 | Casr | NM_013803 | 10 | 19 088.5 (1963) | * |
| 4 | Spink8 | NM_183136 | 8 | 53.9 (19) | * |
| 5 | Pla2g7 | NM_013737 | 7 | 5.3 (3.8) | * |
| 6 | Elovl2 | NM_019423 | 6 | 0.5 (0.3) | |
| 7 | MafB | NM_010658 | 6 | 1 842.7 (542) | * |
| 8 | Irs4 | NM_010572 | 5 | 4 166.0 (958) | * |
| 9 | IL7 | NM_008371 | 4 | 658.7 (32) | * |
| 10 | Nedd4 | NM_010890 | 3 | 210.3 (94) | * |
| 11 | Pax1 | NM_008780 | 3 | 2.5 (2.1) | * |
| 12 | Gcm2 | NM_008104 | 3 | 8.2 (4.6) | * |
| 13 | Angpt2 | NM_007426 | 3 | 3.2 (0.7) | * |
| 14 | Eeflg | NM_026007 | 3 | 0.1 (0.1) | |
| 15 | 2610528KiiRik | NM_175184 | 3 | 0.8 (0.1) | |
| 16 | Gpsn2 | NM_134118 | 3 | 0.3 (0.1) | |
| 17 | Hnrnpf | NM_133834 | 3 | 0.5 (0.1) | |
| 17 genes | 12 genes | ||||
| 2 | |||||
| 1 | Dtprp | NM_010088 | 2 | 0.1 (0.0) | |
| 2 | Pbx1 | NM_008783 | 2 | 1.6 (0.7) | |
| 3 | Atp5a1 | NM_007505 | 2 | 0.4 (0.1) | |
| 4 | Zfp9 | NM_011763 | 2 | 1.1 (0.5) | |
| 50 genes | |||||
| 3 | |||||
| 1 | Ndufv1 | NM_133666 | 1 | 0.1 (0.0) | |
| 2 | Neurod | NM_010894 | 1 | 0.3 (0.1) | |
| 3 | Cks2 | NM_025415 | 1 | 0.3 (0.1) | |
| 4 | Ptpn18 | NM_011206 | 1 | 0.1 (0.0) | |
| 150 genes | |||||
| Total | 217 genes | 12 genes | |||
Subtraction screening was carried out as described in ″Methods.″ Of the 217 genes identified in the screening, 17 that appeared more than twice are categorized in group 1. Groups 2 and 3 consist of genes that appeared twice (50 genes) and once (150 genes), respectively. Ratios of expression levels in the 3PP to expression levels in the 2PA were verified by quantitative RT-PCR, and the values indicate averages and standard errors (SEs) of 3 independent measurements.
Twelve of the 17 group 1 genes were confirmed to be expressed at higher levels in 3PP than in 2PA. None of 4 randomly selected genes from group 2 and 4 randomly selected genes from group 3 showed significantly higher expression in 3PP than in 2PA.