pathscan pcreb  (Cell Signaling Technology Inc)

Journal: International Journal of Molecular Sciences

Article Title: Resveratrol-Loaded Pluronic Micelles Ameliorate Scopolamine-Induced Cognitive Dysfunction Targeting Acetylcholinesterase Activity and Programmed Cell Death

doi: 10.3390/ijms252312777

Figure Lengend Snippet: Effect of pure (RVT) and micellar resveratrol (mRVT) (5 and 10 mg/kg) on BDNF and pCREB protein expression in cortex ( A , C ) and hippocampus ( B , D ) of rats with Sco-induced memory deficit. Each column represents mean ± S.E.M. of 6 animals. Data analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test. Significance versus saline-treated group: ### p < 0.001; significance versus Sco-treated group: *** p < 0.001; significance between RVT and mRVT-treated groups: ƒƒƒ p < 0.001.

Article Snippet: Rat pCREB ELISA kit (cat. no. SL1344Ra) was purchased from Sunlong Biotech Co., Ltd., Hangzhou, China.

Techniques: Expressing, Comparison, Saline

Journal: Biology

Article Title: Neuroprotective Effect of Marrubium vulgare Extract in Scopolamine-Induced Cognitive Impairment in Rats: Behavioral and Biochemical Approaches

doi: 10.3390/biology13060426

Figure Lengend Snippet: Effect of M. vulgare extract on the expression of BDNF and pCREB in the brains of healthy rats and those with scopolamine-induced memory impairment. The concentrations of BDNF and pCREB were assessed using the Enzyme-Linked Immunosorbent Assay (ELISA) in the rat cortex and hippocampus after a 21-day administration of M. vulgare extract. ( A ) BDNF levels in the cortex. ( B ) BDNF levels in the hippocampus. ( C ) pCREB levels in the cortex. ( D ) pCREB levels in the hippocampus. Data are presented as mean values ± SEM ( n = 6 animals per group). Statistical significance is indicated as follows: ## p < 0.01; ### p < 0.001 compared to the saline-treated group; *** p < 0.001 compared to the scopolamine-treated group.

Article Snippet: According to the manufacturer instructions, the following kits were utilized: the Rat BDNF ELISA Kit (cat. no. E-EL-R1235) from Elabscience and the Rat Phospho cAMP response element binding protein (pCREB) ELISA Kit (cat. no. SL1344Ra) from Sunlong Biotech Co., Ltd. (Hangzhou, China) Measurements were performed with a microplate reader set to 450 nm.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Saline

Journal: medRxiv

Article Title: Elevated levels of IL-6 in IgA nephropathy patients are induced by an epigenetically-driven mechanism triggered by viral and bacterial RNA

doi: 10.1101/2022.03.05.22271944

Figure Lengend Snippet: (A) ELISA assay showed significant increased levels of IL-6 in IgAN patients with VTRNA2-1/PKR/CREB activated pathway respect to healthy group. (B) An IL-6 increase was found in T-IgAN patients with VTRNA2-1/PKR/CREB activated pathway compared to TP group, even if not statistically significant. Both the levels of pPKR (C) and pCREB (D) significantly correlated with IL-6 levels in IgAN patients (r=0.97, p= 0.0006 and r=0.89, p=0.0064, respectively). Data are representative of 8 independent experiments (means ±SEM; * p-value < 0.05).

Article Snippet: Colorimetric Cell-Based ELISA assays were used to detect the levels of CREB/pCREB (CREB (Phospho-Ser142) Aviva Systems Biology, USA), and PKR/pPKR (PKR (Phospho-Thr258) Aviva Systems Biology, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Nox1/Ref-1-mediated activation of CREB promotes Gremlin1-driven endothelial cell proliferation and migration

doi: 10.1016/j.redox.2019.101138

Figure Lengend Snippet: Nox1 oxidase mediates hypoxia-induced ROS production, PKA activation, CREB phosphorylation, and CREB: DNA binding in HPAECs. ( A ) HPAECs transfected with scrambled or Nox1 siRNA were exposed to hypoxia for 24 h, and immunoblotting was performed for Nox1 ( n = 5–13); *p < 0.05 and * **p < 0.001. For ( B and C ), HPAECs were exposed to normoxia or hypoxia for 24 h, and hypoxia-treated cells were treated for the final 30 min with 2 µM scrambled (Scrambled) or selective Nox1 peptide inhibitor NoxA1ds. (B ) Evaluation of extracellular O 2 •- was made using HPr + probe ( n = 8–12) * p < 0.05 and * *p < 0.01; and ( C ) cellular H 2 O 2 was determined using CBA probe ( n = 6–10); * *p < 0.01 and * ** *p < 0.0001. For subsequent experiments, HPAECs were transfected with scrambled or Nox1 siRNA and exposed to 24 h hypoxia. ( D ) Analysis of PKA activity by ELISA ( n = 3–5); *p < 0.05. ( E ) Analysis by Western blot of pCREB and tCREB protein levels ( n = 6–13), data show comparisons of pCREB/tCREB ratios; * *p < 0.01. ( F ) Evaluation of CRE-specific DNA binding of pCREB by TransAM © pCREB transcription factor ELISA kit ( n = 3–4); **p < 0.01. All data are shown as means ± S.E.M. Two-way ANOVA followed by Sidak’s post-test was performed for statistical analyses.

Article Snippet: Nuclear extract kit (Cat. 40010) and pCREB TransAM © transcription factor ELISA kit (Cat. 43096) were purchased from Active Motif (Carlsbad, CA, U.S.A.).

Techniques: Activation Assay, Phospho-proteomics, Binding Assay, Transfection, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Nox1/Ref-1-mediated activation of CREB promotes Gremlin1-driven endothelial cell proliferation and migration

doi: 10.1016/j.redox.2019.101138

Figure Lengend Snippet: Hypoxia induces nuclear co-translocation of CREB and Ref-1, nuclear CREB:Ref-1 binding and CREB:DNA-binding in HPAECs. HPAECs were exposed to hypoxia for 24 h. ( A ) Western blot analysis (nuclear lysate) of pCREB ( n = 3); * ** *p < 0.0001, and ( B ) Ref-1 ( n = 6); * **p < 0.001. ( C ) Confocal immunofluorescence images of nuclear pCREB (green staining) ( n = 3); *p < 0.05, and nuclear Ref-1 (red staining) ( n = 3); bar graph shows relative intensity in arbitrary units, *p < 0.05. White staining (DAPI) represents nuclei. ( D ) Analysis of nuclear CREB/Ref-1 binding in HPAECs exposed to hypoxia for 24 h was performed by co-immunoprecipitation (Co-IP). For the Co-IP, anti-pCREB antibody was used for immunoprecipitation, and anti-Ref-1 antibody was used for immunoblotting, and PC= Ref-1 positive control, and Ponceau image was use to experiment normalization ( n = 3–4); *p < 0.05. Nuclear fractions were prepared from HPAECs transfected with scrambled or Ref-1 siRNA exposed to normoxia or hypoxia for 24 h, and ( E ) Western blot for Ref-1 protein levels, ( n = 3–6); *p < 0.05 and * ** *p < 0.0001; and ( F ) CRE-specific DNA binding of pCREB by TransAM © pCREB transcription factor ELISA kit were performed, ( n = 3–4); *p < 0.05. All data are shown as means ± S.E.M. Two-way ANOVA followed by Sidak’s post-test or Student t -test was performed.

Article Snippet: Nuclear extract kit (Cat. 40010) and pCREB TransAM © transcription factor ELISA kit (Cat. 43096) were purchased from Active Motif (Carlsbad, CA, U.S.A.).

Techniques: Translocation Assay, Binding Assay, Western Blot, Immunofluorescence, Staining, Immunoprecipitation, Co-Immunoprecipitation Assay, Positive Control, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Nox1/Ref-1-mediated activation of CREB promotes Gremlin1-driven endothelial cell proliferation and migration

doi: 10.1016/j.redox.2019.101138

Figure Lengend Snippet: Hypoxia induces Nox1-controlled CREB binding to Gremlin1 Cre and CREB- and Ref-1-dependent Gremlin1 expression . ( A ) Analysis of CREB binding at the CRE motif within the human GREM1 promoter in HPAECs exposed to hypoxia for 24 h; pCREB antibody was used for chromatin immunoprecipitation (CHiP); NTC-Non-template control, IgG= rabbit IgG, Nor= Normoxia, Hy= Hypoxia ( n = 2–3). ( B ) Western blot analysis of CREB in HPAECs following transfection with scrambled or CREB siRNA ( n = 3); * **p < 0.001. ( C ) HPAECs transfected with scrambled or CREB siRNA were exposed to normoxia or hypoxia for 24 h, and Western blot was performed for Gremlin1 protein levels, ( n = 5–11); * *p < 0.01 and * ** *p < 0.0001. ( D ) HPAECs transfected with scrambled or Ref-1 siRNA were exposed to normoxia or hypoxia for 24 h, and Western blotting was conducted for Gremlin1 expression, ( n = 3); *p < 0.05; and ( E ) PCNA expression, ( n = 3); *p < 0.05. All data are shown as means ± S.E.M. Two-way ANOVA followed by Sidak’s post-hoc test or Student t -test was performed.

Article Snippet: Nuclear extract kit (Cat. 40010) and pCREB TransAM © transcription factor ELISA kit (Cat. 43096) were purchased from Active Motif (Carlsbad, CA, U.S.A.).

Techniques: Binding Assay, Expressing, Chromatin Immunoprecipitation, Control, Western Blot, Transfection

Journal: Redox Biology

Article Title: Nox1/Ref-1-mediated activation of CREB promotes Gremlin1-driven endothelial cell proliferation and migration

doi: 10.1016/j.redox.2019.101138

Figure Lengend Snippet: Rat PAH is associated with increased Nox1, CREB, CREB:Cre binding and Gremlin1 expression. Rats were treated with a single injection of VEGF receptor (VEGFr) inhibitor SU5416 (20 mg/Kg) followed by 3 weeks in a hypoxia ventilated chamber (10% O 2 ) or were subjected to 3 wks normoxia. VEGFr inhibition exacerbates chronic-hypoxia induced PAH. ( A ) RV maximum pressure, and ( B ) RV hypertrophy as measured by Fulton index (RV/left ventricule+septum weight ratio), ( n = 8); * ** *p < 0.0001. Western blotting of ( C ) Nox1, ( D ) CREB, and ( E ) Gremlin1 protein levels in lung homogenates from Su-Hy-treated rats compared with normoxic controls, ( n = 4); *p < 0.05 and * *p < 0.01. Protein expression was normalized to β-actin protein expression. ( F ) Evaluation of CRE-specific DNA binding of pCREB by TransAM © pCREB transcription factor ELISA kit ( n = 3–4); *p < 0.05. Absorbance (OD at 450 nM) was quantified and expressed as a function of normoxic controls. All data are shown as means ± S.E.M. Student’s t -test was performed.

Article Snippet: Nuclear extract kit (Cat. 40010) and pCREB TransAM © transcription factor ELISA kit (Cat. 43096) were purchased from Active Motif (Carlsbad, CA, U.S.A.).

Techniques: Binding Assay, Expressing, Injection, Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay