ezh2 overexpression plasmid  (Ribobio co)

Journal: eBioMedicine

Article Title: Targeting miR-30d reverses pathological cardiac hypertrophy

doi: 10.1016/j.ebiom.2022.104108

Figure Lengend Snippet: EZH2 regulates H3K27me3 enrichment on the miR-30d promotor. ( a ) Genomic views of the H3K27me3 modification of miR-30d related to transverse aortic constriction (TAC) induced cardiac hypertrophy (GSE93752). ( b ) qRT-PCR analysis of the expression of H3K27 methyltransferase (EZH1 and EZH2) and demethylase (KDM6A and KDM6B) mRNA in neonatal rat cardiomyocytes (NRCMs) treated with phenylephrine (PE, n =6 per group) and angiotensin II (Ang II, n = 6 per group). ( c ) Western blot analysis of the expression of EZH2 protein level in NRCMs treated with PE ( n = 6 per group) and Ang II ( n = 6 per group). ( d ) Dual-luciferase reporter assay showed the regulation between EZH2 and miR-30d in AC16 ( n = 6 per group). ( e ) ChIP assay was performed to detect EZH2 binding and H3K27me3 enrichment on the miR-30d promotor region in AC16 ( n = 3 per group). ( f ) ChIP assay was performed to detect H3K27me3 level enrichment on the miR-30d promotor region when AC16 was treated with EZH2 overexpression plasmid (OE) or negative control Fugw plasmid for 48 h ( n = 3 per group). ( g ) ChIP assay was performed to detect H3K27me3 level enrichment on the miR-30d promotor region when AC16 was treated with EZH2 siRNA or control siRNA ( n = 3 per group). ( h ) Left, Representative immunofluorescence image of a-actinin (red)- and DAPI (blue)-stained NRCM transfected with miR-30d inhibitor and EZH2 siRNA before treated with or without PE for 48 h. Scale bar, 100 μm. Right, Quantification of the average cell surface areas of NRCM transfected with miR-30d inhibitor and EZH2 siRNA before treated with or without PE. ( n = 4 per group, number of CM≥ 50 cells per sample). Data are presented as mean±SD. Statistical significance was determined by Student t test (b-c, e) and two-way ANOVA test with post hoc tukey (d, f-h). *, P < 0.05; **, P < 0.01 and ***, P < 0.001.

Article Snippet: EZH2 overexpression plasmid was purchased from vigenebio (CH882009, China).

Techniques: Modification, Quantitative RT-PCR, Expressing, Western Blot, Luciferase, Reporter Assay, Binding Assay, Over Expression, Plasmid Preparation, Negative Control, Control, Immunofluorescence, Staining, Transfection

Journal: eBioMedicine

Article Title: Targeting miR-30d reverses pathological cardiac hypertrophy

doi: 10.1016/j.ebiom.2022.104108

Figure Lengend Snippet:

Article Snippet: EZH2 overexpression plasmid was purchased from vigenebio (CH882009, China).

Techniques: Recombinant, Western Blot, Lysis, Staining, Software

Journal: Open Medicine

Article Title: miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2

doi: 10.1515/med-2022-0429

Figure Lengend Snippet: Primer sequences used in miRNA reverse transcription and PCR

Article Snippet: The EZH2 overexpressing plasmid was purchased by Ribobio (Guangzhou, China).

Techniques: Reverse Transcription

Journal: Open Medicine

Article Title: miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2

doi: 10.1515/med-2022-0429

Figure Lengend Snippet: miR-22-5p was significantly upregulated in the testicular tissues of patients with cryptorchidism. Testicular tissues of cryptorchidism patients (cry group) and normal testicular tissues of fertile participants (NC group) were collected. (a) The expression of miR-22-5p was detected by QRT-PCR. (b–e) The expression of EZH2 in the testicular tissues was detected by QRT-PCR, western blot, and immunohistochemistry (scale bar = 50 µm). *** p < 0.001 vs NC group.

Article Snippet: The EZH2 overexpressing plasmid was purchased by Ribobio (Guangzhou, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: Open Medicine

Article Title: miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2

doi: 10.1515/med-2022-0429

Figure Lengend Snippet: miR-22-5p directly targets EZH2. (a) The target region of the EZH2 3′UTR for miR-22-5p and the mutant type of EZH2 3′UTR. (b) Effects of miR-22-5p on the activity of firefly luciferase reporters containing either wild-type (Wt) or mutant-type (Mut) 3′UTR were assessed by luciferase reporter gene assays. (c–e) Effects of miR-22-5p on EZH2 expression levels were examined by qRT-PCR and western blot analyses. * p < 0.05, ** p < 0.01 vs mimic NC; ## p < 0.01 vs inhibitor NC.

Article Snippet: The EZH2 overexpressing plasmid was purchased by Ribobio (Guangzhou, China).

Techniques: Mutagenesis, Activity Assay, Luciferase, Expressing, Quantitative RT-PCR, Western Blot

Journal: Open Medicine

Article Title: miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2

doi: 10.1515/med-2022-0429

Figure Lengend Snippet: miR-22-5p regulates SSCs’ self-renewal by targeting EZH2. Human SSCs were co-transfected with miR-22-5p mimics and the EZH2 overexpression plasmid. (a and b) Cell proliferation was measured by EdU staining. (c and d) Cell apoptosis was analyzed by Annexin V/PI staining. (e and f) The expression of SSC markers (GDNF and DAZL) and apoptosis-related proteins (Caspase-3, Bax and Bcl-2) were detected by western blot. * p < 0.05, ** p < 0.01 vs mimic NC + vector; # p < 0.05, ## p < 0.01 vs miR-22-5p mimic + vector.

Article Snippet: The EZH2 overexpressing plasmid was purchased by Ribobio (Guangzhou, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Staining, Expressing, Western Blot