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Results: 5

1.
Figure 4

Figure 4. From: Exaptation of an ancient Alu short interspersed element provides a highly conserved vitamin D-mediated innate immune response in humans and primates.

The human CAMP gene does not respond to retinoic acid or thyroid hormone treatment. The human myeloid cell line NB4 was treated with either vehicle (UNT), 1,25(OH)2D3 (D3), retinoic acid (RA), methoprene acid (MA) or thyroid hormone (T3). Expression of either A) CAMP, B) MPO or C) CDllb was determined by QRT-PCR and normalized to 18S rRNA.

Adrian F Gombart, et al. BMC Genomics. 2009;10:321-321.
2.
Figure 5

Figure 5. From: Exaptation of an ancient Alu short interspersed element provides a highly conserved vitamin D-mediated innate immune response in humans and primates.

The NWM CAMP gene responds to retinoic acid. The marmoset (C. jacchus) B-cell line B95-8 was treated with increasing doses of either ATRA (0.5 to 500 nM) or T3 (0.1 to 100 nM) for 24 h. Relative fold-change compared with untreated cells was determined for CAMP gene expression (normalized to 18S rRNA) using QRT-PCR. CAMP gene expression decreased for ATRA treatment in a dose-dependent fashion. CAMP gene expression was unaffected by T3 treatment.

Adrian F Gombart, et al. BMC Genomics. 2009;10:321-321.
3.
Figure 1

Figure 1. From: Exaptation of an ancient Alu short interspersed element provides a highly conserved vitamin D-mediated innate immune response in humans and primates.

Conservation of the VDRE-containing AluSx SINEs in humans, apes, OWMs and NWMs. A) Amplification of the AluSx SINEs from a panel of non-human primates and humans. Products of the expected size were found in all but three primates. The increase in fragment sizes for M. mulatta, S. labiatus and C. jacchus was due to an additional Alu insertion. B) Schematic indicating the position and type of Alu insertion that was identified from sequencing the PCR products amplified in panel A. The location of the VDRE is indicated by the arrow. C) The nucleotide sequences of each AluSx SINE amplified in panel A were aligned with the AluSx consensus sequence [74]. The positions of the A- and B-boxes are indicated by an underline and the position of the VDRE is outlined by a box.

Adrian F Gombart, et al. BMC Genomics. 2009;10:321-321.
4.
Figure 3

Figure 3. From: Exaptation of an ancient Alu short interspersed element provides a highly conserved vitamin D-mediated innate immune response in humans and primates.

Potential retinoic acid and thyroid hormone receptor binding sites conserved in AluSx of NWMs. The nucleotide sequence (1–89) of the AluSx of each primate was aligned with the consensus AluSx sequence. The location of four direct repeats is indicated within each box with the consensus sequence indicated below each. The potential retinoic acid receptor binding sites (DR2) and the thyroid hormone binding site (DR4) are indicated above the boxes. The locations of the A- and B-boxes of the Alu are indicated below the sequences. The G-to-A and C-to-A change in positions three and five in the third direct repeat occurs in the human, all apes and OWMs, but not NWMs. These changes would likely abrogate binding of TR and RAR to the DR4 and the second DR2, respectively. All NWMs acquired a G-to-A change in the sixth position of the fourth direct repeat thus creating a potentially better DR2 for retinoic acid receptor binding. The third direct repeat in NWMs was unchanged from the consensus Alu sequence and would provide an ideal direct repeat providing a potentially functional DR4 and DR2.

Adrian F Gombart, et al. BMC Genomics. 2009;10:321-321.
5.
Figure 2

Figure 2. From: Exaptation of an ancient Alu short interspersed element provides a highly conserved vitamin D-mediated innate immune response in humans and primates.

Structural and functional conservation of the VDREs in primate promoters. A) The nucleotide sequence of each primate VDRE is aligned to demonstrate the high degree of conservation of the direct repeats (upper case) and 3-bp spacer (lower case). C. aethiops contained a G-to-A change in the second direct repeat otherwise all the direct repeats were identical. B) To determine if primate VDREs were activated by 1,25(OH)2D3, the amplified Alus for H. sapiens, M. mulatta and C. aethiops were subcloned into the pGL4 luciferase reporter vector. The constructs were co-transfected into U937 cells with phTKRL (Promega) to control for efficiency. The change in expression is represented as fold-change comparing vehicle treated to 1,25(OH)2D3 treated cells. C) Mononuclear cells were isolated from the peripheral blood of two individual M. mullata and treated with either vehicle or 100 nM 1,25(OH)2D3 for 48 h. The expression level of CAMP mRNA was determined by QRT-PCR and normalized to 18S levels. The data are represented as ng of CAMP per ng of 18S.

Adrian F Gombart, et al. BMC Genomics. 2009;10:321-321.

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