non phosphorylated forms  (Cell Signaling Technology Inc)

Journal: International journal of molecular sciences

Article Title: Water Extract of Angelica dahurica Inhibits Osteoclast Differentiation and Bone Loss.

doi: 10.3390/ijms241914715

Figure Lengend Snippet: Figure 3. Effects of WEAD on RANK signaling pathways in BMMs. (A) BMMs were treated with or without WEAD (200 µg/mL) and RANKL for the specified durations. The mRNA and protein levels of c-Fos and NFATc1 were assessed by real-time PCR and Western blotting, respectively. (B) BMMs were pre-treated with WEAD or vehicle for 3 h and then stimulated RANKL for the indicated time. Phosphorylated and non-phosphorylated forms of p38, JNK and IκBα were detected using Western blotting. (C) The mRNA expression of Irf8, MafB, Tm7sf4, Atp6v0d2 and CtsK. ** p < 0.01, *** p < 0.001 vs. vehicle control.

Article Snippet: Antibodies against phosphorylated forms including p-p38 (T180/Y182), p-JNK (T183/Y185), p-IκBα (S32) and their non-phosphorylated forms were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Protein-Protein interactions, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Control

Journal: Current Issues in Molecular Biology

Article Title: Network Pharmacological Analysis of a New Herbal Combination Targeting Hyperlipidemia and Efficacy Validation In Vitro

doi: 10.3390/cimb45020086

Figure Lengend Snippet: AT, PC, ZO, and MIX stimulate ACC, AMPK, and AKT phosphorylation in FFA-induced hepatic steatosis and activate CPT-1 related to fatty acid oxidation. HepG2 cells were incubated in the absence or presence of FFAs (1 mM) with AT, PC, ZO, and MIX for 24 h. ( A ) HepG2 cells were treated with various concentrations (0–50 µg/mL) of AT, PC, ZO, and MIX for 24 h. The results are presented as means ± SDs of the percentages determined by three independent experiments versus the non-treated controls. ( B ) HepG2 cells were co-treated with each sample and FFAs for 24 h. Western blot analysis shows the effect on the phosphorylation of ACC and AMPK and AKT protein expression related to the energy metabolism and lipogenesis in FFA-induced hepatic steatosis HepG2 cells. The band intensities were measured by densitometry and normalized versus the intensities of the total forms and β-actin. The results are presented as the means ± SDs of three independent experiments. # p < 0.05 versus FFA-treated controls, and * p < 0.05, ** p < 0.01 versus FFA-treated HepG2 cells.

Article Snippet: The phosphorylation-form or non-phosphorylation-form primary antibodies of ACC (Acetyl-CoA carboxylase), AMPK (AMP-activated protein kinase), AKT (Protein kinase B), CPT-1 (Carnitine palmitoyltransferase-1) were purchased from Cell signaling Technology (Berkeley, CA, USA), and β-actin was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), which also supplied secondary antibodies.

Techniques: Phospho-proteomics, Incubation, Western Blot, Expressing

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study

doi:

Figure Lengend Snippet: Effects of resveratrol (RES) on β-catenin, MnSOD, and PPAR-δ gene expression by real-time PCR. PBMCs were treated with 50 µM RES for 12 h incubation. a) A non-significant change in MnSOD mRNA was observed in healthy subjects after treatment with RES as compared to blank. Treatment with RES could significantly increase the mRNA level of MnSOD in CAD patients compared to blank. b) No significant differences were observed for β-catenin mRNA in healthy subjects and CAD patients after treatment with RES compared to their blank. c) No significant differences in PPAR-δ mRNA were observed for both healthy subjects and CAD patients after treatment with RES compared to their blanks. d) Between-group analysis showed non-significant higher levels of MnSOD, β-catenin and PPAR-δ in healthy subjects than CAD patients in both blank and RES treatment. RES was dissolved in DMSO, and blank groups (untreated cells) were containing only DMSO. In both blank and RES treated cells, DMSO was present at equal concentration (0.025%). All gene expression tests were performed in triplicate in each experiment, and the mean of duplicates were used for statistical analyses. Data are expressed as means ± SEM.

Article Snippet: Total β-catenin protein (non-phosphorylated active form plus phosphorylated inactive form) was measured by enzyme immunoassay (EIA) according to the protocol described in the kit (ADI-900-135; Enzo Life Sciences; USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Incubation, Concentration Assay

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study

doi:

Figure Lengend Snippet: Pearson correlation coefficients of mRNA expression of genes (β-catenin, MnSOD and PPAR-δ). a) In blank groups; β-catenin and MnSOD mRNA expressions were significantly correlated in healthy subjects, while they were not significantly correlated in CAD patients. b) β-catenin and PPAR-δ mRNA showed significant positive correlations in both healthy subjects and CAD patients in blank groups. c) After treatment with resveratrol (RES) a significant positive correlation was found between β-catenin and MnSOD mRNA expressions in healthy subjects, but it was not observed in CAD patients. d) Also β-catenin and PPAR-δ mRNA showed significant positive correlations in both healthy subjects and CAD patients after treatment with RES. RES was dissolved in DMSO, and blank groups (untreated cells) were containing only DMSO. In both blank and RES treated cells, DMSO was present at equal concentration (0.025%).

Article Snippet: Total β-catenin protein (non-phosphorylated active form plus phosphorylated inactive form) was measured by enzyme immunoassay (EIA) according to the protocol described in the kit (ADI-900-135; Enzo Life Sciences; USA).

Techniques: Expressing, Concentration Assay

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study

doi:

Figure Lengend Snippet: Effects of resveratrol (RES) on MnSOD enzyme activity and total β-catenin protein. PBMCs were treated with 50 µM RES for 12 h incubation. a) MnSOD enzyme activity was non-significantly increased in healthy subjects after treatment with RES. Also it was significantly increased in CAD patients after treatment with RES in comparison with blank. b) After treatment with RES a non-significant increasing trend was found for total β-catenin protein of healthy subjects compared to blank. Total β-catenin protein of CAD patients did not significantly increase after treatment with RES. c) Between-group differences showed higher MnSOD enzyme activitiy and total β-catenin protein levels for healthy subjects in comparison with CAD patients. RES was dissolved in DMSO, and blank groups (untreated cells) were containing only DMSO. In both blank and RES treated cells, DMSO was present at equal concentration (0.025%). All experiments were performed in duplicate, and the mean of duplicates were used for statistical analyses. Data are expressed as means ± SEM

Article Snippet: Total β-catenin protein (non-phosphorylated active form plus phosphorylated inactive form) was measured by enzyme immunoassay (EIA) according to the protocol described in the kit (ADI-900-135; Enzo Life Sciences; USA).

Techniques: Activity Assay, Incubation, Concentration Assay

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study

doi:

Figure Lengend Snippet: Pearson correlation coefficients of total β-catenin protein and MnSOD enzyme activity. a) In blank group; total β-catenin protein and MnSOD enzyme activity were significantly correlated in healthy subjects, while they were not significantly correlated in CAD patients. b) After treatment with resveratrol (RES), total β-catenin protein and MnSOD enzyme activity showed a significant positive correlation in healthy subjects, while no significant correlation was observed in CAD patients. RES was dissolved in DMSO, and blank groups (untreated cells) were containing only DMSO. In both blank and RES treated cells, DMSO was present at equal concentration (0.025%)

Article Snippet: Total β-catenin protein (non-phosphorylated active form plus phosphorylated inactive form) was measured by enzyme immunoassay (EIA) according to the protocol described in the kit (ADI-900-135; Enzo Life Sciences; USA).

Techniques: Activity Assay, Concentration Assay

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study

doi:

Figure Lengend Snippet: Effects of resveratrol (RES) on β-Catenin/Wnt and FOXO signaling pathways in PBMCs of CAD patients. The disrupted β-Catenin/FOXO pathway in CAD patients might be due to prolonged exposure to oxidative stress, which exerts a pathogenic role in CAD. RES could increase the MnSOD enzyme activity, however it was not through the β-Catenin/FOXO pathway. The β-catenin/Wnt pathway was intact in CAD patients but was not provoked by RES. (LRP5/6, Low density lipoprotein receptor-related protein 5/6; APC, Adenomatosis polyposis coli; GSK-3β, glycogen synthase kinase-3β; CK1, Casein kinase 1; TCF/LEF, T-cell factor/lymphoid enhancer factor; FOXO, Forkhead box O; MnSOD, Manganese superoxide dismutase; PPAR-δ, Proxisome prolifrator-activated receptor delta

Article Snippet: Total β-catenin protein (non-phosphorylated active form plus phosphorylated inactive form) was measured by enzyme immunoassay (EIA) according to the protocol described in the kit (ADI-900-135; Enzo Life Sciences; USA).

Techniques: Activity Assay

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Dimethyl itaconate protects against lipopolysaccharide-induced endometritis by inhibition of TLR4/NF-κB and activation of Nrf2/HO-1 signaling pathway in mice

doi: 10.22038/ijbms.2020.44151.10346

Figure Lengend Snippet: Effects of DI on LPS-induced NF-κB activation. Mice were administered DI (5, 10, and 20 mg/body weight, IP) 24 hr before LPS induction. The expression of NF-κB p65, NF-κB pp65, IκB, and p-IκB were detected by western blot. β-actin was used as a control. The values presented are mean ± SEM of three parallel measurements. P -value< 0.001:#vs. the control group and P -value< 0.001:***vs. the LPS group. DI: dimethyl itaconate, LPS: lipopolysaccharide, NF-κB: nuclear factor-kappa B

Article Snippet: Primary antibodies for β-actin, toll-like receptor 4 (TLR4), Nrf2, heme oxygenase (HO-1) and phosphorylated and non-phosphorylated forms of nuclear factor-kappa B (NF-κB) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA).

Techniques: Activation Assay, Expressing, Western Blot, Control