Journal: Journal of molecular endocrinology

Article Title: Activation of mitogen-activated protein kinases by a splice variant of GHRH receptor.

doi: 10.1677/JME-09-0121

Figure Lengend Snippet: Figure 6 Activation of ERK1/2 by 1 mM GHRH in HeLa cells transfected with empty vector pcDNA3, pcDNA3-GHRHR and pcDNA3-SV1 plasmids after a period of 0, 5, or 10 min and densitometric analysis of the protein bands. Protein levels were normalized to ERK2 signal (loading control). The blot is representative of two independent experiments with similar results.

Article Snippet: The blots were stripped (Restore Plus Western Blot Stripping Buffer, Thermo Scientific, Waltham, MA, USA) and probed for ERK2 (sc-81457, Santa Cruz Biotechnology) and b-actin (Santa Cruz Biotechnology).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Control

Journal: Cardiovascular research

Article Title: Role of neuronal nitric oxide synthase in lipopolysaccharide-induced tumor necrosis factor-alpha expression in neonatal mouse cardiomyocytes.

doi: 10.1016/j.cardiores.2007.03.020

Figure Lengend Snippet: Fig. 3. Effects of LPS and nNOS on ERK phosphorylation in neonatal mouse cardiomyocytes. Cardiomyocytes from wild-type (WT) and nNOS deficient (nNOS−/−) mice were treated with LPS (10 μg/mL) for 4 h and ERK1/2 phosphorylation was examined by Western blot analysis. Top panel shows a representative blot. Lower panel shows quantitative data expressed as the ratio of phosphorylated ERK1/2 to total ERK1/2. n=7 independent experiments per group. ⁎Pb0.05 vs. corresponding controls, †Pb0.05 vs. WT with LPS.

Article Snippet: Cells were transfected with 100 nM of ERK1 siRNA, scrambled siRNA (Santa Cruz), or equivalent volume vehicle (transfection reagent without siRNA) as described in our previous studies [9,19].

Techniques: Phospho-proteomics, Western Blot

Journal: Cardiovascular research

Article Title: Role of neuronal nitric oxide synthase in lipopolysaccharide-induced tumor necrosis factor-alpha expression in neonatal mouse cardiomyocytes.

doi: 10.1016/j.cardiores.2007.03.020

Figure Lengend Snippet: Fig. 4. Effects of ERK1 siRNA on LPS-induced TNF-α protein expression. A, Western blot demonstrating specific knockdown of ERK1 by ERK1 siRNA. Cardiomyocytes were treated with media (control), scrambled siRNA, or ERK1 siRNA (100 nM) for 24 h. B, Cardiomyocytes were treated with media, scrambled siRNA, vehicle (transfection reagent), or ERK1 siRNA for 24 h prior to LPS administration (10 μg/mL for 4 h). TNF-α protein levels were measured by ELISA and are expressed as the amount of TNF-α protein/100 μL culture medium. n=3–7 independent experiments per group. ⁎Pb0.05 vs. WTcounterpart, †Pb0.05 vs. all other corresponding LPS treated groups.

Article Snippet: Cells were transfected with 100 nM of ERK1 siRNA, scrambled siRNA (Santa Cruz), or equivalent volume vehicle (transfection reagent without siRNA) as described in our previous studies [9,19].

Techniques: Expressing, Western Blot, Knockdown, Control, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Cardiovascular research

Article Title: Role of neuronal nitric oxide synthase in lipopolysaccharide-induced tumor necrosis factor-alpha expression in neonatal mouse cardiomyocytes.

doi: 10.1016/j.cardiores.2007.03.020

Figure Lengend Snippet: Fig. 6. Effects of calcium channel inhibition on LPS-induced TNF-α expres- sion and ERK1/2 phosphorylation. Cultured wild-type (WT) and nNOS−/−

Article Snippet: Cells were transfected with 100 nM of ERK1 siRNA, scrambled siRNA (Santa Cruz), or equivalent volume vehicle (transfection reagent without siRNA) as described in our previous studies [9,19].

Techniques: Inhibition, Phospho-proteomics, Cell Culture

Journal: Frontiers in Cardiovascular Medicine

Article Title: Momordica. charantia -Derived Extracellular Vesicles-Like Nanovesicles Protect Cardiomyocytes Against Radiation Injury via Attenuating DNA Damage and Mitochondria Dysfunction

doi: 10.3389/fcvm.2022.864188

Figure Lengend Snippet: MCELNs ameliorated mitochondrial dysfunction induced cell injury after radiation. (A) H9C2 cells after 48 h of culture with indicated treatment were incubated with mitochondria ROS probe (red) for 15 min. Then nucleus were stained with DAPI (blue) for confocal imaging, scale bar: 20 μm. Flow analysis (B) and quantitation (C) of mitochondria ROS intensity in H9C2 cells after 48 h of culture with indicated treatment. Flow analysis (D) and quantitation (E) of mitochondria membrane potential in H9C2 cells after 48 h of culture with indicated treatment that detected by JC-1 probe. Western blot images (F) and quantitation (G) of AKT, p-AKT, ERK, and p-ERK in H9C2 cells after 48 h of culture with indicated treatment. IR (–/+): 0/16 Gy X-ray; MCELNs (–/+): 0/10 μg/mL. All data were represented as means ± SD ( n = 3 independent experiments). The statistical significance was evaluated by one-way ANOVA followed by the Turkey's multiple comparisons test among groups.

Article Snippet: The primary antibodies included TSG101 (#28283-1-AP, proteintech), CD63 (#25682-1-AP, proteintech), CD9 (#20597-1-AP, proteintech), PCNA (#A0264, ABclonal), Cyclin D1 (#2978, CST), Cyclin B1 (#55004-1-AP, proteintech), cleaved caspase3 (#9664, CST), cleaved PARP (#9548, CST), γ-H2A.x (#ab2893, Abcam), ATM (#NB100-309, NOVUS), p-ATM (#NB100-306, NOVUS), AKT (#60203-2-lg, proteintech), p-AKT (#4060, CST), ERK (#BF8004, affinity), p-ERK (#AF1015, affinity), β-actin (#66009-1-lg, proteintech), α-tubulin (#66031-1-lg, proteintech).

Techniques: Incubation, Staining, Imaging, Quantitation Assay, Membrane, Western Blot