Journal: Journal of Endocrinology

Article Title: CITED2 is expressed in human adrenocortical cells and regulated by basic fibroblast growth factor

doi: 10.1677/joe-06-0083

Figure Lengend Snippet: Figure 1 Representative sections of human embryonic adrenal glands at 8 weeks of gestation. Panel A: negative control without primary antibody (adZadrenal, 100!). Panel B: immunostaining with antibodies to CITED2 revealed that CITED2 is located in the definitive zone (arrows, 100!). Panel C: high magnification of a negative control demonstrates nuclear morphology in the definitive zone (1000!). Panel D: high magnification of a representative section of the definitive zone. Predominantly, small nuclei were stained with an antibody to CITED2 (arrows, 1000!). Panel E: immunohistochemistry with an antibody to 17-a-hydroxylase identified the fetal zone (zF) while the definitive zone (zD) did not stain (100!). Panel F: chromogranin A immunoreactive cells in the outside margin of the adrenal gland (arrows, 100!).

Article Snippet: Then cells were incubated with monoclonal anti-CITED2 antibodies (Novus Biologicals) in a dilution of 1:100 for 1 h at room temperature.

Techniques: Negative Control, Immunostaining, Staining, Immunohistochemistry

Journal: Journal of Endocrinology

Article Title: CITED2 is expressed in human adrenocortical cells and regulated by basic fibroblast growth factor

doi: 10.1677/joe-06-0083

Figure Lengend Snippet: Figure 2 Representative sections of stained normal adult adrenal glands and adrenocortical carcinomas. Panel A: negative control for normal adult adrenal gland without primary antibody (40!). Panel B: normal adult adrenal gland stained for CITED2. Arrows indicate immunoreactivity in the zonae glomerulosa and reticularis at the corticomedullary junction (40!). Panel C: adrenocortical carci- noma stained for CITED2 showed strong nuclear expression of CITED2 (200!). Panel D: gel of RT-PCR analysis of mRNA from normal adult human adrenal cortex (lane 2) and from NCI-H295R cells (lane 3) the 69 bp fragments corresponded to the predicted amplification product of CITED2. As control for PCR we used RNA skipping the reverse transcription reaction (lane 1).

Article Snippet: Then cells were incubated with monoclonal anti-CITED2 antibodies (Novus Biologicals) in a dilution of 1:100 for 1 h at room temperature.

Techniques: Staining, Negative Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Reverse Transcription

Journal: Journal of Endocrinology

Article Title: CITED2 is expressed in human adrenocortical cells and regulated by basic fibroblast growth factor

doi: 10.1677/joe-06-0083

Figure Lengend Snippet: Figure 3 Regulation of CITED2-promotor activity and mRNA-level in NCI-H295R cells. Panel A: relative activity of the CITED2- promotor in NCI-H295R cells after exposure to bFGF in different concentrations either with or without PD98059 for 48 h. Panel B: relative levels of CITED2-mRNA in NCI-H295R cells after exposure to bFGF in different concentrations for 8 h. Panel C: effects of ACTH and forskolin (Forsk) on the CITED2-promotor activity after 48 h of stimulation (left panel) or on the CITED2-mRNA levels (right panel) after 8 h of stimulation in NCI-H295R cells. In all panels P-values indicate significant differences from the control (CNT).

Article Snippet: Then cells were incubated with monoclonal anti-CITED2 antibodies (Novus Biologicals) in a dilution of 1:100 for 1 h at room temperature.

Techniques: Activity Assay, Control

Journal: Journal of Endocrinology

Article Title: CITED2 is expressed in human adrenocortical cells and regulated by basic fibroblast growth factor

doi: 10.1677/joe-06-0083

Figure Lengend Snippet: Figure 4 Immunofluorescence technique demonstrates CITED2 protein expression in NCI-H295R cells. Panel A: unstimulated control. Blue fluorescence dye DAPI marks nuclei. CITED2 protein is represented by a green fluorescence signal in the nuclei. Panel B: exposure of NCI-H295R cells for 24 h to bFGF (10 ng/ml) increased CITED2 nuclear expression in comparison with the control. Panel C: exposure of NCI-H295R cells to bFGF (10 ng/ml) in combination with PD98059 (20 mM) for 24 h attenuated CITED2 nuclear fluorescence signal in comparison with the stimulation with bFGF (10 ng/ml) alone. Panel D: exposure of NCI-H295R cells to ACTH (100 nM) for 24 h did not exert an effect on CITED2-protein expression in comparison with the control.

Article Snippet: Then cells were incubated with monoclonal anti-CITED2 antibodies (Novus Biologicals) in a dilution of 1:100 for 1 h at room temperature.

Techniques: Expressing, Control, Comparison

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The expression levels of stem cell markers importin13, c-kit, CD146, and telomerase are decreased in endometrial polyps

doi: 10.12659/MSM.881901

Figure Lengend Snippet: The expression levels of IPO13 and caspase3 in the endometrial tissues were detected by immunohistochemistry (400×). ( A ) Control endometrium at the proliferation phase. ( B ) Control endometrium at the secretion phase. ( C ) EP at the proliferation phase. ( D ) EP at the secretion phase. ( E ) Control endometrium at the proliferation phase. ( F ) Control endometrium at the secretion phase. ( G ) EP at the proliferation phase. ( H ) EP at the secretion phase. * p<0.05 ( vs. control endometrium at the proliferation phase); ** p<0.05 ( vs. control endometrium at the secretion phase); error bars, SEM.

Article Snippet: Anti-IPO13 goat polyclonal antibody (1:100, NB100-1369, Novus Biologicals), anti-telomerase rabbit polyclonal antibody (1: 100, ZA-0239, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-CD146 mouse monoclonal antibody (1:50, ZM-0299, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-caspase3 rabbit polyclonal antibody (1:100, BS1518, Bioworld Technology, Inc.), anti-bax rabbit polyclonal antibody (1:100, BS1725, Bioworld Technology, Inc.) and anti-bcl-2 rabbit monoclonal antibody (1:100, bs-0522R, Bioworld Technology, Inc.) were incubated with the sections overnight at 4°C.

Techniques: Expressing, Immunohistochemistry, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The expression levels of stem cell markers importin13, c-kit, CD146, and telomerase are decreased in endometrial polyps

doi: 10.12659/MSM.881901

Figure Lengend Snippet: The mRNA expression of IPO13, c-kit, bcl-2 and bax was detected by real-time PCR. (A) Control endometrium at the proliferation phase. (IB) Control endometrium at the secretion phase. (IC) EP at the proliferation phase. (ID) EP at the secretion phase. (IIA) Control endometrium at the proliferation phase. (IIB) Control endometrium at the secretion phase. (IIC) EP at the proliferation phase. (IID) EP at the secretion phase. (IIIA) Control endometrium at the proliferation phase. (IIIB) Control endometrium at the secretion phase. (IIIC) EP at the proliferation phase. (IIID) EP at the secretion phase. (IVA) Control endometrium at the proliferation phase. (IVB) Control endometrium at the secretion phase. (IVC) EP at the proliferation phase. (IVD) EP at the secretion phase. ** p<0.05 ( vs. control endometrium at the secretion phase); error bars, SEM.

Article Snippet: Anti-IPO13 goat polyclonal antibody (1:100, NB100-1369, Novus Biologicals), anti-telomerase rabbit polyclonal antibody (1: 100, ZA-0239, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-CD146 mouse monoclonal antibody (1:50, ZM-0299, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-caspase3 rabbit polyclonal antibody (1:100, BS1518, Bioworld Technology, Inc.), anti-bax rabbit polyclonal antibody (1:100, BS1725, Bioworld Technology, Inc.) and anti-bcl-2 rabbit monoclonal antibody (1:100, bs-0522R, Bioworld Technology, Inc.) were incubated with the sections overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: The expression levels of stem cell markers importin13, c-kit, CD146, and telomerase are decreased in endometrial polyps

doi: 10.12659/MSM.881901

Figure Lengend Snippet: The relative expression levels of IPO13 and bcl-2 proteins in normal endometrial and EP tissues.

Article Snippet: Anti-IPO13 goat polyclonal antibody (1:100, NB100-1369, Novus Biologicals), anti-telomerase rabbit polyclonal antibody (1: 100, ZA-0239, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-CD146 mouse monoclonal antibody (1:50, ZM-0299, Beijing Golden Bridge Biotechnology Co., Ltd.), anti-caspase3 rabbit polyclonal antibody (1:100, BS1518, Bioworld Technology, Inc.), anti-bax rabbit polyclonal antibody (1:100, BS1725, Bioworld Technology, Inc.) and anti-bcl-2 rabbit monoclonal antibody (1:100, bs-0522R, Bioworld Technology, Inc.) were incubated with the sections overnight at 4°C.

Techniques: Expressing, Control

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: The Role of miR-182-5p in Hepatocarcinogenesis of Trichloroethylene in Mice.

doi: 10.1093/toxsci/kfw246

Figure Lengend Snippet: Figure 4. Cited2 is a directly target of miR-182-5p.

Article Snippet: The membrane was immediately immersed in 5% BSA in TBS containing 0.1% Tween-20 for 2 h, and then probed with primary antibodies against Cited2 (Proteintech, Wuhan, China) and b-actin (BBI, Shanghai, China) overnight at 4 C. After washing, blots were labeled with HRP-conjugated secondary anti-rabbit antibody.

Techniques:

Journal: Biochimica et biophysica acta

Article Title: Down-regulation of EBAF in the heart with ventricular septal defects and its regulation by histone acetyltransferase p300 and transcription factors smad2 and cited2.

doi: 10.1016/j.bbadis.2013.07.013

Figure Lengend Snippet: Fig. 7. smad2, cited2, and p300 coordinate to up-regulate the expression of EBAF. (A) cited2, p300, and smad2 cooperatively up-regulated EBAF promoter activity. HCM cells were transfected with EBAF reporter plasmid, together with smad2, p300, and cited2 expression plasmids, and lysed for luciferase reporter assays. (B) smad2, cited2, and p300 were presented in the same complex by Co-IP assays. HCM cells transfected with the smad2-GFP expression plasmid were prepared and precipitated with the anti-smad2 antibody, then detected using Western blotting with anti-p300 or anti-cited2 antibody.

Article Snippet: The proteins were separated on a 12% SDS-PAGE gel and then transferred to polyvinylidene fluoride membrane for Western blotting detection with anti-cited2 (Abcam) or anti-p300 antibody (Santa Cruz).

Techniques: Expressing, Activity Assay, Transfection, Plasmid Preparation, Luciferase, Co-Immunoprecipitation Assay, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Down-regulation of EBAF in the heart with ventricular septal defects and its regulation by histone acetyltransferase p300 and transcription factors smad2 and cited2.

doi: 10.1016/j.bbadis.2013.07.013

Figure Lengend Snippet: Fig. 6. p300 over-expression up-regulates the expression of EBAF by enhancing the acety- lation levels of histone H4 at the EBAF promoter. (A) p300 up-regulated EBAF promoter activity by luciferase reporter assay. (B) Over-expression of p300 increased the enrich- ment of p300 at the EBAF promoter. HCM cells transfected with p300 expression plasmid were subjected to ChIP assays. (C) The acetylation levels of histones H4 were assessed using ChIP assays. HCM cells were transfected with p300 expression plasmid and harvested for ChIP assays. Samples were immunoprecipitated using anti-Ac-H3 or Ac-H4 antibodies, and the precipitated DNAs were amplified using PCR.

Article Snippet: The proteins were separated on a 12% SDS-PAGE gel and then transferred to polyvinylidene fluoride membrane for Western blotting detection with anti-cited2 (Abcam) or anti-p300 antibody (Santa Cruz).

Techniques: Over Expression, Expressing, Activity Assay, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Biochimica et biophysica acta

Article Title: Down-regulation of EBAF in the heart with ventricular septal defects and its regulation by histone acetyltransferase p300 and transcription factors smad2 and cited2.

doi: 10.1016/j.bbadis.2013.07.013

Figure Lengend Snippet: Fig. 8. EBAF down-regulation led to abnormal activation of NODAL-PITX2C pathway. (A) Comparison of PITX2C mRNA expression in 32 samples (16 samples with VSDs and 16 samples without), as revealed by quantitative PCR. **p b 0.01 versus normal group. (B) Comparison of PITX2C mRNA expression between the normal controls (combined) and the different sub- types of VSDs, respectively. M-VSD, S-VSD, A-VSD represent membranous ventricular septal defects, supracristal ventricular septal defects, and atrioventricular canal type defects, respec- tively. **p b 0.01 versus normal group. (C) The up-regulation of PITX2C promoter activity by NODAL overexpression was attenuated when EBAF expression plasmid was co-transfected with NODAL expression plasmid. HCM cells were transfected with EBAF reporter plasmid, together with NODAL and EBAF expression plasmids, and lysed for luciferase reporter assays. *p b 0.05 versus transfection of vector group; **p b 0.01 versus transfection of vector group; #p b 0.05 versus transfection of NODAL expression plasmid only group. Data are from three independent experiments. (D) A sketched diagram shows the regulatory mechanisms driven by smad2, cited2, p300, EBAF and NODAL-PITX2C pathway in heart development.

Article Snippet: The proteins were separated on a 12% SDS-PAGE gel and then transferred to polyvinylidene fluoride membrane for Western blotting detection with anti-cited2 (Abcam) or anti-p300 antibody (Santa Cruz).

Techniques: Activation Assay, Comparison, Expressing, Real-time Polymerase Chain Reaction, Activity Assay, Over Expression, Plasmid Preparation, Transfection, Luciferase