Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: Assessment of H3K27me3 marks in normal and tumoral tissues using ChIP-qPCR. ChIP analysis indicates the change of H3K27me3 marks at six gene loci in prostate cancer tissues. H3K27me3 level on EZH2 was found to be lower in prostate cancer tissues (n = 32) versus normal tissues (n = 33). Contrariwise, the H3K27me3 level of RAR beta 2, PGR , ER alpha and RGMA was significantly higher in tumoral tissues that in normal tissues. The H3K27me3 level on SRC3 in cancer tissue did not reach statistical significance compared in normal tissue. The data is expressed as % of input. Analysis of variance, followed by a Tukey multiple comparison test, was used for statistical analysis. The statistical significant between groups was indicated by letters “a”, “b” and “c”. (N = normal; GS = Gleason score).

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques: Comparison

Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: Factorial Discriminant Analysis (FAD) carried out to describe the variables that discriminate the three groups of patients. RAR beta 2, ER alpha, RGMA and PGR variables conflict to EZH2. Variable SRC3 being away from the circle, it wears a low information (left panel). The graphical representation of patients was used to identify homogeneous groups within the population from the point of view of variable studied (H3K27me3). Red plots represent normal patients (N), green plots represent patients with Gleason score ≤ 7 (<=7) and blue plots represent patients with Gleason score >7 (>7) (right panel).

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques:

Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: EZH2 , AR and SRC3 is upregulated in prostate cancer and inversely correlated with RAR beta 2 , ER alpha , PGR and RGMA . mRNA expression of seven genes were measured using RT-qPCR in normal tissues (n = 38) and cancerous tissues (n = 34). 18S RNA was used as an internal control in PCR reactions. All genes that have high H3K27me3 levels in their promoter are consistent with the low mRNA expression levels. In contrast, genes that have low H3K27me3 levels are consistent with high mRNA expression levels. Analysis of variance, followed by a Tukey multiple comparison test, was used for statistical analysis. The statistical significant between groups was indicated by letters “a”, “b” and “c”. (N = normal; GS = Gleason score).

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques: Expressing, Quantitative RT-PCR, Control, Comparison

Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: Principal component analysis (PCA) carried out to explore the difference between genes and relationships with clinicopathological parameters. The analysis of gene expression was made by PCA. On the dimension 1 (Dim 1), a clear discrimination can be noted between overexpressed genes ( AR , SRC3 and EZH2 ) and under expressed genes ( RAR beta 2 , ER alpha, PGR and RGMA ). The overexpression of AR , SRC3 and EZH2 gene correlated with PSA level (PSA), clinical stage (patho) and Gleason score (Gleason).

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques: Expressing, Over Expression

Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: EZH2 Expression is inversely correlated to RAR beta 2, ER alpha, PGR and RGMA expressions. Protein expressions were analyzed using antibodies against SRC3 (155 KDa), AR (110 KDa), EZH2 (98 KDa), PGR (80 KDa), ER alphax (66KDa), RAR beta 2 (55 KDa) and RGMA (49 KDa). Anti-actin antibody (44 KDa) was used as the internal loading control. Representative data of 3 independent experiments is shown (left panel). Quantification of the western blot data is shown (right panel). The data is normalized to GAPDH (value = means ± SD, all results are statistically significant, *p < 0.05, **p < 0.001). The data is expressed as relative protein level. (N = Normal; T = Tumor).

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques: Expressing, Control, Western Blot

Journal: BMC Cancer

Article Title: The association between Histone 3 Lysine 27 Trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters

doi: 10.1186/1471-2407-14-994

Figure Lengend Snippet: Forward, reverse primers and probes used for ChIP-qPCR for amplification of the gene promoter region

Article Snippet: Primary antibodies used were as follow: anti-EZH2 (1:500, pAb-039-050, Diagenode), anti-RAR beta 2 (1:500, GTX12011, GeneTex), anti-ER alpha (1:250, AC-066-100, Diagenode), anti-PGR (1:250, PAB12723, Abnova), anti-RGMA (1:250, H00056963-M01, Abnova), anti-SRC3 (1:750, MAB7999, Abnova, Walnut, CA, USA), anti-AR (1:1000, GTX73078, GeneTex, Irvine, CA, USA) and anti-β Actin (1 :4000, CP01, Millipore).

Techniques: Amplification