Journal: Oxidative Medicine and Cellular Longevity

Article Title: Depleted Long Noncoding RNA GAS5 Relieves Intervertebral Disc Degeneration via microRNA-17-3p/Ang-2

doi: 10.1155/2022/1792412

Figure Lengend Snippet: Silencing of GAS5 ameliorates IVDD in vivo by miR-17-3p-mediated inhibition of Ang-2. (a) Observation of the MRI signal intensity and IVDD degree by MRI scanning. (b) Western blot analysis of Ang-2 and ECM metabolism-related proteins in NP tissues extracted from IVDD mice, which was normalized to GAPDH. (c) Statistical results of TUNEL-positive cells in the NP tissues extracted from IVDD mice. (d) The expression of glucosaminoglycan (GAG) in mouse NP tissues detected by Safranin O-Fast Green staining. n = 8. (e) HE histological scores of mouse NP tissues by HE staining. n = 8. ∗ p < 0.05 vs. mice without any injection; # p < 0.05 vs. IVDD mice injected with harboring shRNA against shRNA NC; $ p < 0.05 vs. IVDD mice injected with lentivirus harboring shRNA against GAS5; $ p < 0.05 vs. IVDD mice injected with lentivirus harboring overexpression vector NC; @ p < 0.05 vs. IVDD mice injected with lentivirus harboring oe-GAS5. Data (mean ± standard deviation) were assessed by one-way ANOVA, followed by Tukey's post hoc test. n = 8.

Article Snippet: Afterwards, plasmids of sh-NC, sh-GAS5, oe-NC, oe-GAS5, miR-17-3p mimic, and miR-17-3p inhibitor were packaged into lentivirus vectors (pSIH1-H1-copGFP and pLV-EGFP-N) by GenePharma Co., Ltd. (Shanghai, China) to generate lentiviruses including pSIH1-H1-copGFP-sh-NC, pSIH1-H1-copGFP-sh-GAS5, pLV-EGFP-NC, pLV-EGFP-GAS5, pSIH1-H1-copGFP-miR-17-3p inhibitor, and pLV-EGFP-miR-17-3p, respectively.

Techniques: In Vivo, Inhibition, Western Blot, TUNEL Assay, Expressing, Staining, Injection, shRNA, Over Expression, Plasmid Preparation, Standard Deviation

Journal: BMC Cardiovascular Disorders

Article Title: Mycn ameliorates cardiac hypertrophy-induced heart failure in mice by mediating the USP2/JUP/Akt/β-catenin cascade

doi: 10.1186/s12872-024-03748-8

Figure Lengend Snippet: Mycn upregulation alleviates myocardial dysfunction in mice. Mice were administrated with lentiviral vectors-carried OE-Mycn or OE-NC, followed by ISO injection. A , HW/BW of mice. B , Myocardial injury of mice analyzed by HE staining. C , LVEF and LVFS or mice analyzed by echocardiographic assessment. D , Serum levels of CK, CK-MB, LDH, and cTnT in mice determined using ELISA kits. E , ROS content in the mouse heart determined using DHE staining. F - G , MDA content (F) and SOD activity (G) in the mouse heart tissue determined using ELISA kits. H , Concentrations of ANP and BNP in the mouse heart tissue determined using ELISA kits. I , Cell apoptosis in the mouse heart tissue examined using TUNEL assay. J , Mycn mRNA expression in the mouse heart tissue examined by RT-qPCR. In each group, n = 10. Differences were analyzed using the unpaired t test or the two-way ANOVA. * p < 0.05

Article Snippet: For artificial gene interference in mice, the animals were injected with lentiviral vectors-packaged gene overexpression plasmids of Mycn (OE-Mycn), USP2 (OE-USP2), gene knockdown plasmid of JUP (KD-JUP), or the negative control (NC) plasmids (OE-NC and KD-NC) (All provided by VectorBuilder Inc., Guangzhou, Guangdong, China).

Techniques: Injection, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, TUNEL Assay, Expressing, Quantitative RT-PCR

Journal: BMC Cardiovascular Disorders

Article Title: Mycn ameliorates cardiac hypertrophy-induced heart failure in mice by mediating the USP2/JUP/Akt/β-catenin cascade

doi: 10.1186/s12872-024-03748-8

Figure Lengend Snippet: Mycn activates USP2 transcription. A , USP2 predicted as a target of Mycn predicted in the hTFtarget system and the putative binding sites. B - C , mRNA (B) and protein (C) expression of USP2 in the heart tissue of mice after HCM modeling and OE-Mycn treatment examined by RT-qPCR and WB analysis, respectively. D , binding relationship between Mycn and the USP2 promoter in mouse cardiomyocytes determined by the ChIP assay. E , transcription activity of USP2 promoter in mouse cardiomyocytes transfected with OE-Mycn examined by the luciferase reporter gene assay. For animal experiments, n = 10 in each group. For cellular experiments, three biological replicates were conducted. Differences were analyzed by the unpaired t test or the one-way ANOVA. * p < 0.05. See full-length blot images in Supplementary Figure

Article Snippet: For artificial gene interference in mice, the animals were injected with lentiviral vectors-packaged gene overexpression plasmids of Mycn (OE-Mycn), USP2 (OE-USP2), gene knockdown plasmid of JUP (KD-JUP), or the negative control (NC) plasmids (OE-NC and KD-NC) (All provided by VectorBuilder Inc., Guangzhou, Guangdong, China).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Activity Assay, Transfection, Luciferase, Reporter Gene Assay

Journal: BMC Cardiovascular Disorders

Article Title: Mycn ameliorates cardiac hypertrophy-induced heart failure in mice by mediating the USP2/JUP/Akt/β-catenin cascade

doi: 10.1186/s12872-024-03748-8

Figure Lengend Snippet: USP2 overexpression alleviates cardiac hypertrophy and dysfunction in mice. Mice were administrated with lentiviral vectors-carried OE-USP2 or OE-NC, followed by ISO injection. A , HW/BW of mice. B , Myocardial injury of mice analyzed by HE staining. C , LVEF and LVFS or mice analyzed by echocardiographic assessment. D , Serum levels of CK, CK-MB, LDH, and cTnT in mice determined using ELISA kits. E , ROS content in the mouse heart determined using DHE staining. F , SOD activity and MDA content in the mouse heart tissue determined using ELISA kits. G , Concentrations of ANP and BNP in the mouse heart tissue determined using ELISA kits. H , Cell apoptosis in the mouse heart tissue examined using TUNEL assay. I , Mycn mRNA expression in the mouse heart tissue examined by RT-qPCR. In each group, n = 10. Differences were analyzed using the unpaired t test or the two-way ANOVA. * p < 0.05

Article Snippet: For artificial gene interference in mice, the animals were injected with lentiviral vectors-packaged gene overexpression plasmids of Mycn (OE-Mycn), USP2 (OE-USP2), gene knockdown plasmid of JUP (KD-JUP), or the negative control (NC) plasmids (OE-NC and KD-NC) (All provided by VectorBuilder Inc., Guangzhou, Guangdong, China).

Techniques: Over Expression, Injection, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, TUNEL Assay, Expressing, Quantitative RT-PCR

Journal: BMC Cardiovascular Disorders

Article Title: Mycn ameliorates cardiac hypertrophy-induced heart failure in mice by mediating the USP2/JUP/Akt/β-catenin cascade

doi: 10.1186/s12872-024-03748-8

Figure Lengend Snippet: Graphical abstract. In the context of cardiac hypertrophy, Mycn ameliorates heart failure by activating transcription of USP2, which enhances the stability of JUP protein via deubiquitination modification and subsequently blocks the Akt/β-catenin pathway

Article Snippet: For artificial gene interference in mice, the animals were injected with lentiviral vectors-packaged gene overexpression plasmids of Mycn (OE-Mycn), USP2 (OE-USP2), gene knockdown plasmid of JUP (KD-JUP), or the negative control (NC) plasmids (OE-NC and KD-NC) (All provided by VectorBuilder Inc., Guangzhou, Guangdong, China).

Techniques: Modification