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Results: 6

1.
FIGURE 6.

FIGURE 6. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

Reduction in selenium levels in tissues of GPx1 KO mice. Selenium was determined by ICP-MS in tissues from wild type (WT) and GPx1 knock-out (KO) mice. Values are means ± S.D. (error bars). Organs in which selenium was analyzed are shown below each panel (**, p < 0.005; *, p < 0.05).

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.
2.
FIGURE 3.

FIGURE 3. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

Transcriptome analysis and characterization of the MR selenoproteome. A, length distribution of assembled contigs of the MR liver transcriptome. B, selenoproteins identified in MR. The human selenoproteome is used as a reference to represent the MR selenoproteome. Selenoproteins identified by sequencing the MR liver transcriptome are highlighted. The location of Sec residues is indicated by a red line. Seven Sec residues were found in SelP. GPx1 has an early termination codon (see “Results”). C, schematic representation of MR GPx1 in comparison with mouse GPx1.

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.
3.
FIGURE 2.

FIGURE 2. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

MR tissue extracts have low GPx1 expression and activity. Total MsrB (A), MsrA (B), and GPx (C) activities were measured in the indicated mouse and MR tissues. D, primary splenocytes derived from MR and mice were metabolically labeled with 75Se, and protein extracts were analyzed by SDS-PAGE followed by autoradiography. The band corresponding to GPx1 is shown with an arrow on the right. E, relative expression of GPx1 mRNA in MR and mouse liver (*, p < 0.05). Error bars, S.D.

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.
4.
FIGURE 1.

FIGURE 1. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

XFM and ICP-MS analysis of selenium in mouse and MR tissues. Shown are XFM scans of mouse and MR livers (A) and testes (B). Each element and its maximum and minimum threshold values are given above each image in ng/cm2. The rainbow-colored scale bar relates to the signal intensity measured as ng/cm2 in each spot, with dark pixels representing areas of low concentration and a gradient to bright pixels depicting increasing concentrations. A scale bar is shown below the elemental maps. C, selenium was analyzed by ICP-MS in mouse and MR tissues. Values are means ± S.D. (error bars). Organs in which trace elements were analyzed are shown below each panel.

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.
5.
FIGURE 5.

FIGURE 5. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

SECIS element does not decrease GPx1 expression level. A, alignment of mammalian GPx1 SECIS elements. Nucleotides critical for the SECIS function are underlined. B, MR GPx1 SECIS element as predicted by SECISearch. Functional sites are shown in boldface type (24). C, mouse GPx1 SECIS element. Functional sites are shown in boldface type. D, mouse and MR GPx1 coding sequences were cloned into pSelExpress1 containing an efficient eukaryotic SECIS element and expressed in HEK 293 cells. Cells were labeled with 75Se, followed by SDS-PAGE, autoradiography, and Western blotting with Myc antibodies. Coomassie staining is a loading control. E, coding sequences of mouse GPx1 containing the MR GPx1 3′-UTR (M-long-MR SECIS) and coding sequence of MR GPx1 containing mouse GPx1 3′-UTR (MR-short-M SECIS) were expressed in HEK 293 cells. Cells were labeled with 75Se, followed by SDS-PAGE, autoradiography, and Western blotting with Myc and GFP antibodies. WB, Western blot.

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.
6.
FIGURE 4.

FIGURE 4. From: Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression.

GPx1 is poorly expressed in mammalian cells. A, schematic representation of MR and mouse GPx1 mutant constructs. Positions of Sec and glutamine (CAG) codons, the stop signal (TAG), and the SECIS element in the 3′-UTR are shown. B, mRNA levels were assessed by real-time PCR and normalized to GFP expressed from the same vector. Results are given ± S.D. (error bars). C, MR and mouse Myc-GPx1 constructs were transfected into HEK 293 cells. Cells were labeled with 75Se, followed by SDS-PAGE and autoradiography (top). Migration of ectopically expressed GPx1 is shown on the left. The same lysates were probed with anti-Myc and anti-β-actin antibodies (two bottom panels). Myc-GPx1 mutants were also immunoprecipitated with anti-Myc antibodies, followed by PhosphorImager analysis of the 75Se-labeled GPx1. D, mouse and MR constructs were transfected into HEK 293 cells followed by treatment of cells with 10 μm MG132 for 12 h. Cells were labeled with 75Se, followed by protein analysis by SDS-PAGE and autoradiography (top). The same membrane was stained with Myc and GFP antibodies. E, expression of mouse and MR cysteine mutants of GPx1 in HEK 293 cells was analyzed by Western blotting. IP, immunoprecipitation; WB, Western blotting.

Marina V. Kasaikina, et al. J Biol Chem. 2011 May 13;286(19):17005-17014.

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