Journal: Journal of neurochemistry

Article Title: Bone morphogenetic protein 2 inhibits neurite outgrowth of motor neuron-like NSC-34 cells and up-regulates its type II receptor.

doi: 10.1111/j.1471-4159.2012.07795.x

Figure Lengend Snippet: Fig. 5 Mash1 and Id1 expression during differentiation of NSC-34 cells. Total protein samples from undifferentiated (0) or differentiated NSC-34 cells were analyzed by western blot using anti-Mash1 (a) or anti-Id1 (b) antibodies. a-tubulin was used as loading control. Bars represent the Mean ± SD values obtained from band intensity analy- ses of three independent experiments (*p < 0.05; **p < 0.01). (c) NSC-34 cells were transfected with an Id1 promoter-luciferase re- porter gene (pId1-Luc). Enzyme activity was measured before or after 24 and 48 h of differentiation and normalized by Renilla luciferase activity (pRL-SV40). Mean ± SD of three independent experiments performed in triplicate are shown (***p < 0.001).

Article Snippet: Antibodies against a-tubulin (Sigma-Aldrich, St. Louis, MO, USA), Mash1 (BD Pharmingen, San Diego, CA, USA), Id1, BMPR-II, BMPR-IA and BMPR-IB (Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho-p38 MAPK, phospho-Smad 1, phospho-LIMK1, phospho-AKT and LIMK1 (Cell Signaling Technologies, Frankfurt, Germany) were used for immunoblotting.

Techniques: Expressing, Western Blot, Control, Transfection, Luciferase, Activity Assay

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: Effect of cyclin D1 and Id1 on breast cancer cell protein expression and migration . Actively cycling MDA-MB-231 and ZR75-1 cells were monitored 20 h post-transfection with the indicated siRNA (cyclin D1/CDK4/6/Id1) or vector (Id1) for changes in gene or protein expression, and migration. Blots are representative, and plots are mean values from at least three independent experiments. Error bars represent standard deviation. MDA-MB-231 cells: (A) Microarray analysis. Left panel: CCND1 gene expression, right panel: ID1 gene expression (B) Western blot for cyclin d1, Id1 and Actin protein, (C) Cell migration as measured by Boyden chamber assay, dots indicate total number of migrated cells. (D) ChIP assay for Id1 promoter region following cyclin D1 pull down. ZR75-1: (E) Western blot for cyclin d1 and Actin protein (F) Cell migration- Boyden chamber assay. ***P ≥ 0.001, **P ≥ 0.01, *P ≥ 0.05 vs. control, two-tailed student's t-test.

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Expressing, Migration, Transfection, Plasmid Preparation, Standard Deviation, Microarray, Gene Expression, Western Blot, Boyden Chamber Assay, Control, Two Tailed Test

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: Effect of cyclin d1 and Id1 on EMT markers . MDA-MB-231 cells were monitored 20 h post-transfection with the indicated siRNA (cyclin D1/CDK4/6/Id1) or vector (Id1) for changes in EMT-related gene expression by microarray analysis. Additionally, MDA-MB-231 and ZR75-1 gene expression was examined by qPCR assay. Plots are mean values from at least three independent experiments Error bars represent standard deviation. (A) Microarray analysis of SNAI1 , SNAI2 , CDH11 , TWIST1 and VIM gene expression. (B-D) CCND1, ID1 and SNAI2 in MDA-MB-231 cells. (E, F) qPCR analysis of CCND1 and SNAI2 in ZR75-1 cells. ***P ≥ 0.001, **P ≥ 0.01, *P ≥ 0.05 vs. control, two-tailed student's t-test.

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Transfection, Plasmid Preparation, Gene Expression, Microarray, Standard Deviation, Control, Two Tailed Test

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: Distribution of CCND1 and ID1 gene expression according to clinico- pathological parameters in breast cancer patients

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Gene Expression

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: Correlation of CCND1 , ID1 , SNAI1 and SNAI2 expression to recurrence free survival . Expression of our genes of interest in relation to recurrence-free survival was examined in a breast cancer database containing 1,107 tumours from Sims et al. (2008). Gene expression intensity was quartiled as 1-low, 2- medium low, 3- medium high and 4- high, and assessed in all patients, ER-positive and ER-negative patients, respectively (A) CCND1 quartiles (B) ID1 quartiles (C) SNAI1 quartiles (D) SNAI2 quartiles. P-value is based on log-rank test.

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Expressing, Gene Expression

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: EMT-related gene expression intensity and recurrence free survival in CCND1 / ID1 high and low tumours . A breast cancer database was employed to examine (A) Mean-centered average expression of EMT-related genes of interest in CCND1 / ID1 subgroups, and (B, C) Recurrence-free survival of ER-positive and negative patients in CCND1 / ID1 subgroups. P-value is based on log-rank test.

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Gene Expression, Expressing

Journal: BMC Cancer

Article Title: Cyclin D1, Id1 and EMT in breast cancer

doi: 10.1186/1471-2407-11-417

Figure Lengend Snippet: Gene expression of Cyclin d1/Id1 and EMT markers in breast cancer cell lines and claudin-low tumours . Expression of EMT and related genes from the Neve et al cell line (A-C) and Herschkowitz et al. claudin-low (D-F) studies. (A and D) Heatmaps showing relative expression of genes of interest, Red = high, Green = low. (B and E) Scatterplots showing the relationship between breast cancer subgroups and CCND1 / ID1 expression (C and F) Proportion of CCND1- low/ ID1 -high cell lines and tumours in breast cancer subgroups. ***P ≥ 0.001 , Chi 2 test.

Article Snippet: 3 μg of anti-cyclin d1 antibody (DCS-6, DAKO, Denmark) was used to pull down cyclin D1, with subsequent detection of Id1 using SimpleChIPTM Human Id1 Promoter Primers (Cell Signalling Technology, Danvers, MA) and Mrg1 as positive control (Eurofins Laboratories Ltd., Manchester, UK).

Techniques: Gene Expression, Expressing