Journal: Molecular Medicine Reports

Article Title: Inhibitor of DNA binding 1 regulates cell cycle progression of endothelial progenitor cells through induction of Wnt2 expression

doi: 10.3892/mmr.2016.5491

Figure Lengend Snippet: Reverse transcription-polymerase chain reaction primer sequences.

Article Snippet: The primary antibodies were as follows: Rabbit anti-mouse Id1 (1:1,000; catalog no. ab134163; Abcam), rabbit anti-mouse cyclin D1 (1:2,000; catalog no. ab134175; Abcam), rabbit anti-mouse Wnt2 (1:500; catalog no. ab27794; Abcam), rabbit anti-mouse β-catenin (1:1,000; catalog no. ab6302; Abcam), rat anti-mouse β-actin (1:1,000; catalog no. aa128; Beyotime Institute of Biotechnology) and rabbit anti-mouse histone (1:1,000; catalog no. ab1791; Abcam).

Techniques: Reverse Transcription Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Inhibitor of DNA binding 1 regulates cell cycle progression of endothelial progenitor cells through induction of Wnt2 expression

doi: 10.3892/mmr.2016.5491

Figure Lengend Snippet: Effects of Id1 on cell cycle progression of EPCs. The cell cycle distribution of EPCs, transfected with (A) Ad-Id1 or (B) si-Id1, was analyzed by flow cytometry. Ad-Id1 transfection decreased the percentage of EPCs in G 1 phase and increased the percentage in S/G 2 M phases, while si-Id1 transfection induced the opposite effect. Cyclin D1 mRNA and protein expression levels from EPCs treated with (C) Ad-Id1 or (D) si-Id1 were detected using reverse transcription-polymerase chain reaction and western blot analysis, respectively. Ad-1d1 transfection increased, and si-Id1 transfection decreased, cyclin D1 mRNA and protein expression levels. The expression level was analyzed relative to β-actin (n=3). * P<0.05 vs. wild type. EPCs, endothelial progenitor cellAd, adenoviruId1, inhibitor of DNA binding 1; si, small interfering; con, control.

Article Snippet: The primary antibodies were as follows: Rabbit anti-mouse Id1 (1:1,000; catalog no. ab134163; Abcam), rabbit anti-mouse cyclin D1 (1:2,000; catalog no. ab134175; Abcam), rabbit anti-mouse Wnt2 (1:500; catalog no. ab27794; Abcam), rabbit anti-mouse β-catenin (1:1,000; catalog no. ab6302; Abcam), rat anti-mouse β-actin (1:1,000; catalog no. aa128; Beyotime Institute of Biotechnology) and rabbit anti-mouse histone (1:1,000; catalog no. ab1791; Abcam).

Techniques: Transfection, Flow Cytometry, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Inhibitor of DNA binding 1 regulates cell cycle progression of endothelial progenitor cells through induction of Wnt2 expression

doi: 10.3892/mmr.2016.5491

Figure Lengend Snippet: Wnt2 knockdown attenuates the effects of Id1 on cell cycle progression in EPCs. (A) The cell cycle distribution of Ad-Id1/si-Wnt2 EPCs was analyzed by flow cytometry. Transfection of Ad-Id1 and si-Wnt2 increased the percentage of EPCs in G 1 phase and decreased the percentage in S/G 2 M phases, compared with Ad-Id1/si-con. (B) The mRNA expression levels of Wnt2, Id1 and cyclin D1 in EPCs were detected by reverse transcription-polymerase chain reaction. (C) The protein expression levels of Wnt2, Id1 and cyclin D1 in EPCs were detected by western blot analysis. The expression levels were analyzed relative to β-actin (n=3). Cyclin D1 mRNA and protein levels were decreased in Ad-Id1/si-Wnt2 EPCs compared with EPCs transfected with Ad-Id1/si-con. * P<0.05 vs. Ad-vector/si-con; # P<0.05 vs. Ad-Id1/si-con. Wnt2, wingless-type mouse mammary tumor virus integration site family member 2; EPCs, endothelial progenitor cellAd, adenoviruId1, inhibitor of DNA binding 1; si, small interfering; con, control.

Article Snippet: The primary antibodies were as follows: Rabbit anti-mouse Id1 (1:1,000; catalog no. ab134163; Abcam), rabbit anti-mouse cyclin D1 (1:2,000; catalog no. ab134175; Abcam), rabbit anti-mouse Wnt2 (1:500; catalog no. ab27794; Abcam), rabbit anti-mouse β-catenin (1:1,000; catalog no. ab6302; Abcam), rat anti-mouse β-actin (1:1,000; catalog no. aa128; Beyotime Institute of Biotechnology) and rabbit anti-mouse histone (1:1,000; catalog no. ab1791; Abcam).

Techniques: Flow Cytometry, Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Plasmid Preparation, Binding Assay

Journal: Journal of Xenobiotics

Article Title: Chronic Exposure to Both Electronic and Conventional Cigarettes Alters Ileum and Colon Turnover, Immune Function, and Barrier Integrity in Mice

doi: 10.3390/jox14030053

Figure Lengend Snippet: Effects of chronic electronic and conventional cigarette exposure on ileum- and colon-proliferation signalling pathways. Transcript levels of markers of major proliferation pathways in ileum ( A ) and colon ( B ). n = 14 per group. Transcript levels of cyclin D1 pathway genes in ileum ( C ) and colon ( D ). n = 14 per group. Cyclin D1 immunostaining, with higher magnification on the right, and quantification of cyclin D1 mean optical density in ileum ( E ) and colon ( F ). Cyclin D1 stain: brown. Nuclear stain: blue. n = 5 per group. * p < 0.05, ** p < 0.01, *** p < 0.005 and **** p < 0.001 compared to the control group (Air), as determined by the Mann–Whitney U test.

Article Snippet: Rabbit anti-mouse Ki-67 primary antibody (MA5-14520, Invitrogen; Fisher Scientific, Illkirch, France 1/100 dilution), rabbit anti-mouse cyclin D1 primary antibody (#55506, Cell Signaling Technology; Danvers, USA 1/400 dilution), rabbit anti-mouse ZO-1 primary antibody (#61-7300, Invitrogen; 1/200 dilution), and rabbit anti-mouse occludin primary antibody (#91131, Cell Signaling; 1/200 dilution) were incubated overnight at 4 °C.

Techniques: Immunostaining, Staining, Control, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: A Short-Term Model of Colitis-Associated Colorectal Cancer That Suggests Initial Tumor Development and the Characteristics of Cancer Stem Cells

doi: 10.3390/ijms241411697

Figure Lengend Snippet: Immunofluorescence staining for ( A ) β-catenin (green), ( B ) Sox9 (red), ( C ) Ki67 (green), ( D ) cyclin D1 (red), and ( E ) CDK4 (green) in the micro-neoplastic lesions of the AOM/DSS mice. DAPI (blue) was used for nuclear staining. ( A – E ) In the right panels, the same sections were stained with hematoxylin and eosin after immunofluorescence staining and observed by light microscopy. Original magnification, ×200 ( A – E ). Scale bars: 300 μm ( A – E ).

Article Snippet: The primary Abs used in this study were as follows: mouse monoclonal anti-human β-catenin Ab (sc-7963, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rat monoclonal anti-mouse Ki67 Ab (652402, BioLegend, San Diego, CA, USA), mouse monoclonal anti-human CDK4 Ab (sc-23896, Santa Cruz Biotechnology), rabbit monoclonal anti-human Sox9 Ab (ab185230, Abcam, Cambridge, UK), goat polyclonal anti-human B cell-specific Moloney murine leukemia virus integration site 1 (Bmi1) Ab (ab115251, Abcam), rabbit monoclonal anti-mouse cyclin D1 Ab (ab16663, Abcam), and rabbit polyclonal anti-human pSmad2/3L-Thr Ab.

Techniques: Immunofluorescence, Staining, Light Microscopy

Journal: International Journal of Molecular Sciences

Article Title: A Short-Term Model of Colitis-Associated Colorectal Cancer That Suggests Initial Tumor Development and the Characteristics of Cancer Stem Cells

doi: 10.3390/ijms241411697

Figure Lengend Snippet: Immunofluorescence staining for ( A ) β-catenin (green), ( B ) Sox9 (red), ( C ) Ki67 (green), ( D ) cyclin D1 (red), and ( E ) CDK4 (green) in the uni-cryptal neoplastic lesions of the AOM/DSS mice. DAPI (blue) was used for nuclear staining. ( A – E ) In the right panels, the same sections were stained with hematoxylin and eosin after immunofluorescence staining and observed via light microscopy. Original magnification, ×200 ( A – E ). Scale bars: 100 μm ( A – E ).

Article Snippet: The primary Abs used in this study were as follows: mouse monoclonal anti-human β-catenin Ab (sc-7963, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rat monoclonal anti-mouse Ki67 Ab (652402, BioLegend, San Diego, CA, USA), mouse monoclonal anti-human CDK4 Ab (sc-23896, Santa Cruz Biotechnology), rabbit monoclonal anti-human Sox9 Ab (ab185230, Abcam, Cambridge, UK), goat polyclonal anti-human B cell-specific Moloney murine leukemia virus integration site 1 (Bmi1) Ab (ab115251, Abcam), rabbit monoclonal anti-mouse cyclin D1 Ab (ab16663, Abcam), and rabbit polyclonal anti-human pSmad2/3L-Thr Ab.

Techniques: Immunofluorescence, Staining, Light Microscopy