pax2 cdna  (New England Biolabs)

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: PAX2 expression level in stable over-expression or knocking-down cell lines. (A) High malignant endometrial carcinoma cell lines exhibited higher PAX2 mRNA expression than cell lines with low malignant potential: AN3CA>HEC1B>HEC1A>RL952. (B) PAX2-si3 had the best interference effect among 3 PAX2 siRNAs in mRNA level. (C) The protein interference effect of PAX2-si3 was obvious in HEC1A and HEC1B. The expression level of PAX2 in HEC1A was 1. (D) The mRNA and protein level of HEC1B-siPAX2 were lower than HEC1B and HEC1B-siNonTarget. (E) The mRNA and protein level of HEC1A-pCMV-PAX2 were higher than HEC1A and HEC1A-pCMV-neo. ** p <0.0001; * p <0.05.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Expressing, Over Expression

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: The difference of biological ability of HEC1A, HEC1A-pCMV-neo and HEC1A-pCMV-PAX2. (A) Viability of HEC1A-pCMV-PAX2 was significantly higher than HEC1B and HEC1B-siNonTarget after 48 hours. HEC1A-pCMV-PAX2 migration ability (B) and nvasion ability (C) were significantly higher than that of the negative control groups. (D) HEC1A-pCMV-PAX2 exhibited obvious accumulation in the S phase population in cell cycle. ** p <0.0001; * p <0.05.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Migration, Negative Control

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: PAX2 promoted endometrial cancer cells progression in xenograft nude mice model. (A) HEC1A, HEC1A-pCMV-neo and HEC1A-pCMV-PAX2 growth situation in nude mice. (B) PAX2 expression level in nude mice subcutaneous tumor. (C) HEC1A-pCMV-PAX2 cells proliferated faster than HEC1A and HEC1A-pCMV-neo cells in vivo. (D) Final weight of HEC1A-pCMV-PAX2 was about 3-times of control groups. (E) HE stain of normal liver tissue and livers with metastatic lesions. (F) HEC1A-pCMV-PAX2 group exhibited visible metastatic lesions of the liver. ** p <0.0001; * p <0.05.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Expressing, In Vivo, H&E Stain

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: Down-regulation PAX2 inhibited endometrial cancer cells progression in xenograft nude mice model. (A) HEC1B, HEC1B-siNonTarget and HEC1B-siPAX2 growth situation in nude mice. (B) PAX2 expression level in nude mice subcutaneous tumor. (C) HEC1B-siPAX2 proliferated more faster than HEC1B and HEC1B-siNonTarget.(D) Final weight of HEC1B-siPAX2 was lighter than control groups. ** p <0.0001; * p <0.05.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: Network generation of CDK1, YWHAZ and PRKDC based on significantly differential genes found between cells with PAX2 over-expression and those that lack that over-expression. Red nodes represented up-regulated genes, and blue ones were down-regulated genes.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Over Expression

Journal: Journal of Cancer

Article Title: Paired box 2 promotes progression of endometrial cancer via regulating cell cycle pathway

doi: 10.7150/jca.22418

Figure Lengend Snippet: PAX2 activated the cell cycle pathway via regulating CDK1. (A) The PAX2 binding site in promoter region of CDK1. (B) The PAX2 binding sites in promoter region of YWHAZ. (C) CDK1 and YWHAZ mRNA level were elevated in HEC1A-pCMV-PAX2.(D)The protein expression level of CDK1 and PAX2 in cells. (E) Comparison of cell viability in HEC1A-pCMV-neo, HEC1A-pCMV-PAX2, HEC1A-pCMV-PAX2 non-targeting and HEC1A-pCMV-PAX2 siCDK1 cells.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV- Myc-neo) were offered by Origene (Rockville, MD, USA).

Techniques: Binding Assay, Expressing

Journal: PLoS Genetics

Article Title: MITF – A controls branching morphogenesis and nephron endowment

doi: 10.1371/journal.pgen.1007093

Figure Lengend Snippet: A) In situ hybridization of Bmp7 , Pax2 and Wnt9b in wild-type (WT) and homozygous (HO) MITF-A transgenic kidneys at E13.5 (magnification X200, n = 5–6 per genotype). B) Quantitative RT-PCR analysis of Bmp7 , Pax2 and Wnt9b mRNA expression in E13.5 kidneys of WT, heterozygous (HE) and HO MITF-A transgenic embryos (n = 6–9 per genotype). C) In situ hybridization of Re t, Wnt11 and Spry1 in WT and HO MITF-A transgenic kidneys at E13.5 (magnification X200, n = 5–6 per genotype). Note the increased staining of Re t mRNA in transgenic kidneys at E13.5. D) Quantitative RT-PCR analysis of Re t, Wnt11 and Spry1 mRNA expression in E13.5 kidneys of WT, HE and HO MITF-A transgenic embryos (n = 6–9 per genotype). Data are means ± SEM. ANOVA followed by Tukey-Kramer test; transgenic versus wild-type mice: * P < 0.05, ** P < 0.01.

Article Snippet: The following cDNA templates were used: Ret , Wnt11 , Pax2 , BMP7 , Wnt9b and Spry1 (a kind gift from Isabelle Gross, INSERM U682).

Techniques: In Situ Hybridization, Transgenic Assay, Quantitative RT-PCR, Expressing, Staining

Journal: Oncotarget

Article Title: DNA methylation promotes paired box 2 expression via myeloid zinc finger 1 in endometrial cancer

doi: 10.18632/oncotarget.12626

Figure Lengend Snippet: A. PAX2 expression was increased in the epithelium of endometrial cancers compared with endometrial hyperplasia and normal tissues. a. type 1 endometrial cancer, b. type 2 endometrial cancer, c. complex hyperplasia, d. atypical hyperplasia, e. proliferative endometrium, f. atrophic endometrium. All images were obtained at 200x magnification using light microscopy. B. PAX2 expression in the epithelium was semi-quantified according to IHC results and compared between different groups. C. PAX2 expression in the stroma was semi-quantified according to IHC results and compared between different groups. **p<0.01, *p<0.05

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV-Neo) were obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Light Microscopy

Journal: Oncotarget

Article Title: DNA methylation promotes paired box 2 expression via myeloid zinc finger 1 in endometrial cancer

doi: 10.18632/oncotarget.12626

Figure Lengend Snippet: A. PAX2 was knocked down or overexpressed by transfection of siRNA or pCMV-Myc-PAX2, respectively, in HEC-1A, HEC-1B and RL95-2 cells. B. Cell proliferation was reduced by knocking down PAX2 and stimulated by overexpressing PAX2 in HEC-1A, HEC-1B and RL95-2 cells. C. Cell migration was decreased in cells transfected with siRNA-PAX2 and increased in cells transfected with pCMV-Myc-PAX2. D. Cell invasion was inhibited in cells transfected with siRNA-PAX2 and stimulated in cells transfected with pCMV-Myc-PAX2. *p<0.05, **p<0.01.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV-Neo) were obtained from Origene (Rockville, MD, USA).

Techniques: Transfection, Migration

Journal: Oncotarget

Article Title: DNA methylation promotes paired box 2 expression via myeloid zinc finger 1 in endometrial cancer

doi: 10.18632/oncotarget.12626

Figure Lengend Snippet: A. Bisulfite sequencing results showed fragment B1 (-764 to -447 bp) of the PAX2 promoter was hypermethylated in endometrial cancer cell lines and tissues: rows represent clones (10 for each sample), columns represent CpG sites. Black squares represent methylated CpGs, and white squares represent unmethylated CpGs. EEC: endometrial epithelial cell. EnCa: endometrial cancer. N: normal. B. MassARRAY results indicate that PAX2 was hypermethylated in endometrial cancer tissues compared with normal endometrial tissues. M1 is an amplicon of a 280-bp fragment from -723 bp to -443 bp; M2 is an amplicon of a 379-bp fragment from -468 bp to -89 bp. Hypermethylated CpG sites were centralized in the 5’ of M1 (CpG 1 to 7).

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV-Neo) were obtained from Origene (Rockville, MD, USA).

Techniques: Methylation Sequencing, Clone Assay, Methylation, Amplification

Journal: Oncotarget

Article Title: DNA methylation promotes paired box 2 expression via myeloid zinc finger 1 in endometrial cancer

doi: 10.18632/oncotarget.12626

Figure Lengend Snippet: A. PAX2 weak expression group (IHC score=0 or 1) exhibited reduced methylation levels compared with the strong expression group (IHC score=2 or 3). B. PAX2 promoter (fragment B1) of endometrial cancer cell lines was demethylated by 5-Aza-CdR treatment. C. PAX2 was down-regulated on mRNA level after 5-Aza-CdR treatment in RL95-2 and HEC-1B cells. *p<0.05. PAX2 expression in EEC1 was reduced compared with endometrial cancer cells. D. PAX2 protein expression was down-regulated after 5-Aza-CdR treatment in Ishikawa, RL95-2 and HEC-1B cells as measured by western blot. E. PAX2 protein was down-regulated after 5-Aza-CdR treatment in endometrial cancer cells measured by immunocytochemistry: a. HEC-1A control, b. HEC-1A 5-Aza-CdR, c. Ishikawa control, d. Ishikawa 5-Aza-CdR, e. RL95-2 control, f. RL95-2 5-Aza-CdR, g. HEC-1B control, h. HEC-1B 5-Aza-CdR. *p<0.05, # p<0.01.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV-Neo) were obtained from Origene (Rockville, MD, USA).

Techniques: Expressing, Methylation, Western Blot, Immunocytochemistry, Control

Journal: Oncotarget

Article Title: DNA methylation promotes paired box 2 expression via myeloid zinc finger 1 in endometrial cancer

doi: 10.18632/oncotarget.12626

Figure Lengend Snippet: A. Deletion analysis of the PAX2 promoter in HEC-1A cells. B. Deletion analysis of the PAX2 promoter in HEC-1B cells. C. MZF1 mRNA was down-regulated in HEC-1B cells 24 to 48 hours after transfection of MZF1-siRNA. D. Luciferase reporter gene activity was increased after knocking down MZF1 in HEC-1B cells transfected with pGL3-P1, but not up-regulated in cells transfected with pGL3-P3, which did not contain the repressive transcriptional region of PAX2 promoter (-683 bp to -393 bp). E. PAX2 protein was up-regulated in HEC-1B cells 72 hours after MZF1-siRNA transfection. F. By analyzing the RNAseq data from TCGA database, an negative correlation was demonstrated between the expression of MZF1 and PAX2 (R=-0.263, p=0.043). *p<0.05.

Article Snippet: Full-length PAX2 cDNA (pCMV-Myc-PAX2) clone and vector (pCMV-Neo) were obtained from Origene (Rockville, MD, USA).

Techniques: Transfection, Luciferase, Activity Assay, Expressing

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Mutations in PAX2 Associate with Adult-Onset FSGS

doi: 10.1681/ASN.2013070686

Figure Lengend Snippet: Table 1.

Article Snippet: 39 , 40 DNA Constructs Full-length HA-tagged PAX2 and Myc-DDK tagged TLE4 cDNAs were provided by OriGene Technologies.

Techniques: