human crbp1 complementary deoxyribonucleic acid (gene number nm-002899)  (OriGene)

Journal: Journal of Diabetes Investigation

Article Title: Electronegative low‐density lipoprotein of patients with metabolic syndrome induces pathogenesis of aorta through disruption of the stimulated by retinoic acid 6 cascade

doi: 10.1111/jdi.13158

Figure Lengend Snippet: Stimulated by retinoic acid 6 (STRA6) cascades decrease and markers of atherosclerosis increase in the aortas of electronegative low‐density lipoprotein (L5)‐injected mice. The protein of aorta samples ( n = 3) was extracted from saline‐injected (control [Ctl]), L1‐injected (L1) and L5‐injected (L5) mice, as well as L5‐injected LOX1 −/− mice (LOX1 −/− +L5) after injection for 4 weeks. (a) Western blots showed that L5 decreased aortic STRA6, cellular retinol‐binding protein 1 (CRBP1), lecithin‐retinol acyltransferase (LRAT), retinoic acid receptor (RAR)α and retinoid X receptor (RXR)α, but increased aortic LOX1, p‐p38 mitogen‐activated protein kinases (p‐p38), pSmad2, transforming grown factor‐β 1 (TGFβ 1 ), caspase 3 (Casp3) and matrix metallopeptidase 9 (MMP9) in L5‐injected mice. In LOX1 −/− mice, these changes caused by L5 were ameliorated. (b) Bar graphs showed that LOX1 increased, and STRA6, CRBP1, LRAT, RARα and RXRα decreased in L5‐injected mice. These changes caused by L5 were recovered in L5‐injected LOX1 −/− mice. (c) Bar graphs show that the p‐p38/p38 and pSmad2/Smad2 ratio increased in L5‐injected mice. These changes were reversed in L5‐injected LOX1 −/− mice. (d) TGFβ 1 , caspase 3 and MMP9 levels of L5‐injected mice increased. These changes were also recovered in L5‐injected LOX1 −/− mice. All results are presented as the mean ± standard error. * P < 0.05 versus Ctl and L1; # P < 0.05 versus L5. WT, wild‐type.

Article Snippet: The plasmid DNA of cytomegalovirus 6‐green fluorescent protein (pCMV6‐GFP) vector and human crbp1 complementary deoxyribonucleic acid (gene number NM‐002899) were purchased from OriGene Technologies Inc. (Rockville, MD, USA).

Techniques: Injection, Saline, Control, Western Blot, Binding Assay

Journal: Journal of Diabetes Investigation

Article Title: Electronegative low‐density lipoprotein of patients with metabolic syndrome induces pathogenesis of aorta through disruption of the stimulated by retinoic acid 6 cascade

doi: 10.1111/jdi.13158

Figure Lengend Snippet: Electronegative low‐density lipoprotein (L5) suppresses stimulated by retinoic acid 6 (STRA6) cascades and induces markers of atherosclerosis in human aortic endothelial cell lines (HAECs). (a) Western blots showed the expression of LOX1, STRA6, cellular retinol‐binding protein 1 (CRBP1), lecithin‐retinol acyltransferase (LRAT), retinoic acid receptor (RAR)α, retinoid X receptor (RXR)α, p‐p38 mitogen‐activated protein kinases (p‐p38), pSmad2, and matrix metallopeptidase 9 (MMP9) in control small interfering ribonucleic acid (siC)‐transfected and LOX1 small interfering ribonucleic acid (siLOX1)‐transfected HAECs ( n = 3) after phosphate‐buffered saline (control [Ctl]), L1 or L5 treatment for 24 h. In L5‐treated HAECs, the quantitative analysis showed that (b) LOX1 protein level significantly increased; but (c) protein levels of STRA6, (d) CRBP1, (e) LRAT, (f) RARα and (g) RXRα decreased; (h) p‐p38/p38 and (i) pSmad2/Smad2 ratios increased; and (j) protein level of MMP9 increased. These changes were reversed in LOX1 small interfering ribonucleic acid‐transfected HAECs under L5 treatment. All results are presented as the mean ± standard error. * P < 0.05 versus Ctl‐ and L1‐treated siC group; # P < 0.05 versus L5‐treated siC group.

Article Snippet: The plasmid DNA of cytomegalovirus 6‐green fluorescent protein (pCMV6‐GFP) vector and human crbp1 complementary deoxyribonucleic acid (gene number NM‐002899) were purchased from OriGene Technologies Inc. (Rockville, MD, USA).

Techniques: Western Blot, Expressing, Binding Assay, Control, Transfection, Saline

Journal: Journal of Diabetes Investigation

Article Title: Electronegative low‐density lipoprotein of patients with metabolic syndrome induces pathogenesis of aorta through disruption of the stimulated by retinoic acid 6 cascade

doi: 10.1111/jdi.13158

Figure Lengend Snippet: Electronegative low‐density lipoprotein (L5) suppresses stimulated by retinoic acid 6 (STRA6) cascades and enhances markers of atherosclerosis in human aortic smooth muscle cell lines (HASMCs). (a) Western blots showed LOX1, STRA6, cellular retinol‐binding protein 1 (CRBP1), lecithin‐retinol acyltransferase (LRAT), retinoic acid receptor (RAR)α, retinoid X receptor (RXR)α, p‐p38 mitogen‐activated protein kinases (p‐p38), p‐Smad2 and matrix metallopeptidase 9 (MMP9) protein expression in control small interfering ribonucleic acid (siC)‐transfected and LOX1 small interfering ribonucleic acid (siLOX1)‐transfected HASMCs ( n = 3) after phosphate‐buffered saline (control [Ctl]), L1 or L5 treatment for 24 h. Quantitative analysis showed that L5 treatment (b) increased LOX1, (c) decreased STRA6, (d) CRBP1, (e) LRAT, (f) RARα and (g) RXRα protein levels, but (h) increased p‐38/p38 and (i) pSmad2/Smad2 ratios, and (j) MMP9 protein level as compared to phosphate‐buffered saline‐ and L1‐treated cells. All these effects of L5 in siC‐transfected cells were reversed in LOX1 small interfering ribonucleic acid‐transfected HASMCs under L5 stimulation. All results were presented as the mean ± standard error. * P < 0.05 versus Ctl‐ and L1‐treated siC group; # P < 0.05 versus L5‐treated siC group.

Article Snippet: The plasmid DNA of cytomegalovirus 6‐green fluorescent protein (pCMV6‐GFP) vector and human crbp1 complementary deoxyribonucleic acid (gene number NM‐002899) were purchased from OriGene Technologies Inc. (Rockville, MD, USA).

Techniques: Western Blot, Binding Assay, Expressing, Control, Transfection, Saline

Journal: Journal of Diabetes Investigation

Article Title: Electronegative low‐density lipoprotein of patients with metabolic syndrome induces pathogenesis of aorta through disruption of the stimulated by retinoic acid 6 cascade

doi: 10.1111/jdi.13158

Figure Lengend Snippet: The crbp1 gene transfection reverses electronegative low‐density lipoprotein (L5) effects on stimulated by retinoic acid 6 (STRA6) cascades and markers of atherosclerosis in human aortic endothelial cell lines (HAECs). (a) Western blots showed LOX1, STRA6, cellular retinol‐binding protein 1 (CRBP1), lecithin‐retinol acyltransferase (LRAT), retinoic acid receptor (RAR)α, retinoid X receptor (RXR), p38 mitogen‐activated protein kinases (p‐38), pSmad2 and matrix metallopeptidase 9 (MMP9) expression in cell lysate of pCMV6‐transfected and pCMV6‐crbp1‐transfected HAECs under phosphate‐buffered saline (control [Ctl]), L1 and L5 treatment for 24 h ( n = 3). Quantitative analysis showed significant differences for (b) the increase of LOX1; the decrease of (c) STRA6, (d) CRBP1, (e) LRAT, (f) RARα and (g) RXRα; the increase of (h) p‐p38/p38 and (i) pSmad2/Smad2 ratios, and (j) MMP9 under L5 stimulation in pCMV6‐transfected cells. These changes caused by L5 treatment were reversed in pCMV6‐crbp1‐transfected cells. All results are presented as the mean ± standard error. * P < 0.05 versus Ctl‐ and L1‐treated plasmid DNA of cytomegalovirus 6 (pCMV6) groups; # P < 0.05 versus L5‐treated pCMV6 group.

Article Snippet: The plasmid DNA of cytomegalovirus 6‐green fluorescent protein (pCMV6‐GFP) vector and human crbp1 complementary deoxyribonucleic acid (gene number NM‐002899) were purchased from OriGene Technologies Inc. (Rockville, MD, USA).

Techniques: Transfection, Western Blot, Binding Assay, Expressing, Saline, Control, Plasmid Preparation

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: L5 decreases STRA6 cascades and injures the kidney. The protein of kidney samples (n = 3) was extracted from saline-injected (Ctl), L1-injected (L1), and L5-injected (L5) mice, as well as L5-injected LOX1 −/− mice (LOX1 −/− +L5) after injection for 4 weeks. A: Western blots showed that L5 reduced STRA6, CRBP1, RARα, RARγ, and RXRα, but increased LOX1, pJNK, p-p38MAPK (p-p38), TGFβ 1 , pSmad2, caspase 3 (Casp3), and collagen 1 (Col1) in L5-injected mice. In LOX1 −/− mice, these changes caused by L5 were diminished. B: Bar graphs show that LOX1 increased and STRA6, CRBP1, RARα, RARγ, and RXRα decreased in L5-injected mice. These changes caused by L5 were attenuated in L5-injected LOX1 −/− mice. C: Bar graphs indicate that pJNK/JNK, p-p38/p38, and pSmad2/Smad2 ratios increased in L5-injected mice. These changes caused by L5 were attenuated in L5-injected LOX1 −/− mice. D: TGFβ 1 , caspase 3, and collagen 1 levels of L5-injected mice increased. These changes caused by L5 were recovered in L5-injected LOX1 −/− mice. E: RT-PCR analysis shows that L5 decreased PCR products of STRA6, CRBP1, RARα, and RXRα in L5-injected mice, but there was no effect in L5-injected LOX1 −/− mice. All results are represented as mean ± SE; * P < 0.05 versus Ctl and L1; # P < 0.05 versus L5.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: Injection, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: L5 suppresses STRA6 cascades and increases cell injury in HK-2 cells. A: Western blots show LOX1, STRA6, CRBP1, RARα, RARγ, RXRα, pJNK, pSmad2, and collagen 1 (Col1) in control siRNA (siC)-transfected or LOX1 siRNA (siLOX1)-transfected HK-2 cells (n = 3) after PBS (Ctl), L1, or L5 treatment for 24 h. In L5-treated HK-2 cells, quantitative analysis showed that LOX1 (B) protein levels significantly increased; STRA6 (C), CRBP1 (D), RARα (E), RARγ (F), and RXRα (G) protein levels decreased; pJNK/JNK (H) and pSmad2/Smad2 (I) ratios increased; and collagen 1 (J) increased. These changes were reversed in LOX1 siRNA-transfected L5-injected HK-2 cells, as compared with L5-injected siC mice. K: Representative images (400×) show immunoreactive staining for STRA6 (red fluorescence) emerged to cell membrane staining (green fluorescence) on chamber slides of Ctl-, L1-, and L5-treated cells, and L5-treated LOX1 siRNA-transfected cells. L: ELISA showed an increase of TGFβ 1 concentration in L5-treated siC cells, but not in L5-treated siLOX1 cells. M: Cytochemistry images show the nucleus of apoptotic cells (green fluorescence) emerged to staining of nucleus (blue fluorescence) in Ctl-, L1-, and L5-treated cells, and in L5-treated LOX1 siRNA-transfected cells. N: Number of apoptotic cells significantly increased in L5-treated siC cells, but not in L5-treated siLOX1 cells. O: Real-time PCR analysis showed that LOX1 siRNA significantly increased mRNA levels of STRA6, CRBP1, RARα, and RXRα in L5-treated HK-2 cells. All results are represented as mean ± SE. * P < 0.05 versus Ctl- and L1-siC; # P < 0.05 versus L5-siC group.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: Western Blot, Transfection, Injection, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: JNK inhibitor represses L5 effect on STRA6 cascades and renal injury. A: Western blots show STRA6, CRBP1, RARα, RARγ, RXRα, pSmad2, and collagen 1 (Col1) of Ctl, L1-, or L5-treated RETCs with or without SP600125 treatment for 24 h (n = 3). Quantitative analysis showed that L5 treatment decreased STRA6 (B), CRBP1 (C), RARα (D), RARγ (E), and RXRα (F), but increased the pSmad2/Smad2 ratio (G) and collagen 1 (H). All of these effects of L5 were reversed in SP600125-treated RETCs under L5-stimulation. I: Representative optical microscopy images show nuclei of apoptotic cells (thick dark stain) in Ctl, L1-treated, and L5-treated cells with or without SP600125 treatment. SP600125 decreased the nuclei of apoptotic cells in RETCs under L5-stimulation. J: ELISA showed that SP600125 treatment attenuated the increase of TGFβ 1 concentration in the culture medium of L5-treated RETCs. K: Real-time PCR analysis revealed that SP600125 significantly recovered the decreased mRNA levels of STRA6, CRBP1, RARα, and RXRα in L5-treated RETCs. All results are represented as mean ± SE. * P < 0.05, ** P < 0.01 versus Ctl and L1; # P < 0.05, ## P < 0.05 versus L5.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: Western Blot, Microscopy, Staining, Enzyme-linked Immunosorbent Assay, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: The crbp1 gene transfection reverses L5 effects on STRA6 cascades and renal cell injury. A: Western blots show LOX1, STRA6, CRBP1, RARα, RXRα, pJNK, pSmad2, and collagen 1 (Col1) expression in cell lysate of pCMV6-transfected and pCMV6-crbp1-transfected HK-2 cells under PBS (Ctl), L1, or L5 treatment for 24 h (n = 3). Quantitative analysis showed a significant difference for the increase of LOX1/actin (B); the decrease of STRA6/actin (C), CRBP1/actin (D), RARα/actin (E), and RXRα/actin (F); the increase of pJNK/JNK (G), pSmad2/Smad2 (H), caspase 3/actin (I), and collagen 1/actin (J) under L5 stimulation in pCMV6-transfected cells. These changes caused by L5 treatment were reversed in pCMV6-crbp1-transfected cells. K: ELISA showed that pCMV6-crbp1-transfection reversed the elevation of TGFβ 1 concentration under L5 treatment in the culture medium of HK-2 cells. L: Cytochemistry images show immunoreactive staining of STRA6 (green fluorescence) was recovered by pCMV6-crbp1-transfection in L5-treated HK-2 cells (crbp1+L5). All results are represented as mean ± SE. § P < 0.05 versus Ctl- and L1-treated pCMV6 groups; * P < 0.05 versus Ctl-treated pCMV6 group; # P < 0.05 versus L1-treated pCMV6 group; + P < 0.05 versus L5-treated pCMV6 group.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Fluorescence

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: STRA6 siRNA damages STRA6 cascades and induces renal cell injury. A: Western blots show LOX1, STRA6, CRBP1, RARα, RXRα, pJNK, pSmad2, and collagen 1 (Col1) in cell lysate of STRA6 siRNA (siSTRA6) or control siRNA (siC) transfected HK-2 cells cultured in PBS (Ctl)-, L1-, and L5-treated HK-2 cells for 24 h (n = 3). Quantitative analysis showed significance for the increase of LOX1/actin (B); the decrease of STRA6/actin (C), CRBP1/actin (D), RARα/actin (E), RXRα/actin (F), pJNK/JNK (G), pSmad2/Smad2 (H), and collagen 1/actin (I) in STRA6 siRNA-transfected HK-2 cells in Ctl and L1 groups. J: ELISA also showed a significant increase of TGFβ 1 in culture medium of siSTRA6-transfected HK-2 cells in Ctl, L1, and L5 groups. K: Cytochemistry images show that STRA6 siRNA transfection increased the nuclei of apoptotic cells (green fluorescence) in Ctl- and L1-treated cells. All results are represented as mean ± SE. § P < 0.05 versus and L1-treated siC cells; * P < 0.05 versus Ctl-treated siC cells; # P < 0.05 versus L1-treated siC cells; + P < 0.05 versus L5-treated siC cells.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: Western Blot, Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Journal of Lipid Research

Article Title: Electronegative low density lipoprotein induces renal apoptosis and fibrosis: STRA6 signaling involved [S]

doi: 10.1194/jlr.M067215

Figure Lengend Snippet: The schematic mechanism of L5 on STRA6 cascade suppression and kidney injury. L5-activated LOX1/JNK and p38MAPK pathway suppresses STRA6/CRBP1/retinol/RA/RARs/RXRs cascade and, thereafter, contributes to apoptosis and fibrosis of the kidney.

Article Snippet: The pCMV6-GFP vector and human crbp1 cDNA (gene number NM_002899) were purchased from OriGene Technologies Inc. (Rockville, MD).

Techniques: