phosphorylated ampk α1  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: Metformin prevents stroke damage in non-diabetic female mice with chronic kidney disease

doi: 10.1038/s41598-021-86905-9

Figure Lengend Snippet: Metformin pre-conditioning enhances adenosine monophosphate-activated protein kinase (AMPK) activation and reduces canonical NFκB activation in CKD mice. ( A ) Representative images of phosho-AMPK, IΚBα, phospho-P65, phospho-ACC (Ser-79) and total-ACC western blots performed on SHAM and CKD mice exposed, or not, to metformin. ( B ) Quantitative data showing higher AMPK phosphorylation in ischemic hemispheres of CKD-metformin than CKD-vehicle treated mice. ( C ) Quantitative data showing higher ACC phosphorylation in ischemic hemispheres of CKD-metformin than CKD-vehicle treated mice. ( D ) Quantitative data showing higher IΚBα expression in ischemic hemispheres of CKD-metformin than CKD-vehicle treated mice. ( E ) Quantitative data showing lower P65 phosphorylation in ischemic hemispheres of CKD-metformin than CKD-vehicle treated mice. Results are expressed as the median, interquartile, and min–max and show data from at least 6 animals per group. Statistical analysis was performed using a non-parametric Mann–Whitney test. *p < 0.05, **p < 0.01, CKD-vehicle vs. SHAM-vehicle mice. $ p < 0.05, $$ p < 0.01, CKD-metformin vs. CKD-vehicle mice.

Article Snippet: Western blotting was performed using rabbit polyclonal anti-phosphorylated AMPK α1/2 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal anti-phosphorylated ACC (ser-79) (1:1000, Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-ACC (1:1000, Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-phosphorylated NFκB p65 (1:1000, Cell Signaling Technology, Danvers, MA), rabbit polyclonal anti-IƘBα (1:1000, Cell Signaling Technology, Danvers, MA), or rabbit polyclonal anti-LC3A/B (1:1000, Cell Signaling Technology, Danvers, MA) as described previously .

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Expressing, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Metformin prevents stroke damage in non-diabetic female mice with chronic kidney disease

doi: 10.1038/s41598-021-86905-9

Figure Lengend Snippet: Hypothetical scheme of the mechanisms by which metformin prevent ischemic stroke damage in non-diabetic mice with CKD. In brain ischemic lesions of non-diabetic CKD mice, AMPK activity, which is known to block microglia/macrophages M1 polarization, is impaired. The recruited microglia/macrophages consequently display increased M1 and reduced M2 polarization. The subsequent inflammation may be responsible for the elevated apoptosis, increased infarct volumes and poorer functional outcomes. A 5-week course of pre-conditioning with metformin rescued the activation of AMPK in mice with CKD. This effect is associated with a significant decrease of macrophage/microglia transition towards the M 1 phenotype, as shown by the decreased expression of the M 1 markers CD16, CD32, and CD86. Metformin-induced AMPK activation blocks NFκB activation and the subsequent release of pro-inflammatory cytokines such as IL-1β. This decrease in inflammation may be responsible for less brain-cell apoptosis, thus reducing the cortical infarct volume and subsequent neurobehavioral disorders.

Article Snippet: Western blotting was performed using rabbit polyclonal anti-phosphorylated AMPK α1/2 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal anti-phosphorylated ACC (ser-79) (1:1000, Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-ACC (1:1000, Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-phosphorylated NFκB p65 (1:1000, Cell Signaling Technology, Danvers, MA), rabbit polyclonal anti-IƘBα (1:1000, Cell Signaling Technology, Danvers, MA), or rabbit polyclonal anti-LC3A/B (1:1000, Cell Signaling Technology, Danvers, MA) as described previously .

Techniques: Activity Assay, Blocking Assay, Functional Assay, Activation Assay, Expressing

Journal: Scientific Reports

Article Title: Cellular and molecular mechanisms associated with ischemic stroke severity in female mice with chronic kidney disease

doi: 10.1038/s41598-019-42933-0

Figure Lengend Snippet: CKD impairs adenosine monophosphate-activated protein kinase (AMPK) activation and promotes canonical NFκB activation. ( A and B ) Study of AMPK activation. ( A ) Representative images of the western blot analysis. ( B ) Quantitative data showing lower AMPK phosphorylation within ischemic hemispheres of CKD mice as compared to SHAM-operated mice. ( C and D ) Study of IκBα degradation. ( C ) Representative images of the western blot analysis. ( D ) Quantitative data showing lower IκBα expression within ischemic hemispheres of CKD mice as compared to SHAM-operated mice. ( E and F ) Study of p65 phosphorylation. ( E ) Representative images of the western blot analysis. ( F ) Quantitative data showing higher phosphorylation of P65 within ischemic hemispheres of CKD mice as compared to SHAM-operated mice. Results are expressed as mean ± SEM and represent data from 4 animals per group. *p < 0.05 CKD versus SHAM mice (non parametric Mann-Whitney U test). Ipsi: ipsilateral hemisphere. Raw western blot data are provided in Supplementary Figs , and .

Article Snippet: Western blotting was performed using rabbit polyclonal anti-phosphorylated AMPK α1/2 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit monoclonal anti-phosphorylated NFƘB p65 (1:1000, Cell Signaling Technology, Danvers, MA) or rabbit polyclonal anti-IƘBα (1:1000, Cell Signaling Technology, Danvers, MA) as detailed in Supplementary Methods.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Expressing, MANN-WHITNEY

Journal: Journal of neuroscience research

Article Title: Hindbrain A2 Noradrenergic Neuron Adenosine 5′-Monophosphate-Activated Protein Kinase (AMPK) Activation, Upstream Kinase/Phosphorylase Protein Expression, and Receptivity to Hormone and Fuel Reporters of Short-Term Food Deprivation are Regulated by Estradiol

doi: 10.1002/jnr.23892

Figure Lengend Snippet: Description of Primary Antibodies Used

Article Snippet: ; rabbit; sc-25792; Lot no. 1009; AB_216946 0.4 ug/mL pAMPK Phosphorylated Thr 172 of human AMPK α1/2 Santa Cruz Biotechnol.

Techniques: Concentration Assay

Journal: Journal of neuroscience research

Article Title: Hindbrain A2 Noradrenergic Neuron Adenosine 5′-Monophosphate-Activated Protein Kinase (AMPK) Activation, Upstream Kinase/Phosphorylase Protein Expression, and Receptivity to Hormone and Fuel Reporters of Short-Term Food Deprivation are Regulated by Estradiol

doi: 10.1002/jnr.23892

Figure Lengend Snippet: Individual tyrosine hydroxylase (TH)-immunoreactive (-ir) A2 neurons were laser-microdissected from 10 μm-thick sections of the caudal DVC of FD and FF groups of E and O rats. For each protein of interest, lysates of pooled A2 cells (n=50 per treatment group; n=10 per rat) were analyzed by Western blot in triplicate at minimum; protein band optical densities (O.D.) were quantified with Syngene Genetool 4.01 software and expressed relative to α-tubulin. Panels depict mean normalized O.D. values ± S.E.M. for A2 neuron AMPK (Figure 2.A) and pAMPK (Figure 2.B) proteins for the following treatment groups: FD/O (white bars), FD/E (gray bars), FF/O (diagonal-striped white bars), FF/E (diagonal-striped gray bars). Representative immunoblots of individual proteins of each interest and α-tubulin are shown below each graph. *p < 0.05, FD versus FF; **p < 0.05, E versus O.

Article Snippet: pAMPK , Phosphorylated Thr 172 of human AMPK α1/2 , Santa Cruz Biotechnol.; rabbit; sc-33524; Lot no. J0915; AB_216714 , 0.4 ug/mL.

Techniques: Western Blot, Software

Journal: Journal of neuroscience research

Article Title: Hindbrain A2 Noradrenergic Neuron Adenosine 5′-Monophosphate-Activated Protein Kinase (AMPK) Activation, Upstream Kinase/Phosphorylase Protein Expression, and Receptivity to Hormone and Fuel Reporters of Short-Term Food Deprivation are Regulated by Estradiol

doi: 10.1002/jnr.23892

Figure Lengend Snippet: At the top of the Figure, side-by-side illustrative depictions of the systemic vasculature system list blood substrate fuel and hormone surrogates of metabolic imbalance potentially reactive to FD accompanied by brackets containing arrows denoting direction of change (↑: increase; ↓: decrease) or symbolic portrayal of absence of change (△ covered by an X) in the presence (left-hand side) versus absence (right-hand side) of estradiol. In the lower half of the figure, representative A2 noradrenergic neurons illustrate FD effects on glucocorticoid receptor (GR) and FFA transporter (ACSVL3) protein content in A2 cells harvested from rats implanted with estradiol (left) or oil (right). The box associated with each A2 neuron shows FD effects on AMPK upstream kinase and phosphorylase, total and phosphorylated AMPK, and pAMPK/AMPK ratio. Below each box brackets indicate effect or lack of effect of FD on A2 DβH protein expression. Abbreviations: ACSVL3, very long-chain acyl-CoA synthetase 3 (also referred to as fatty acid transport protein 3 or FATP3); AMPK, 5′-monophosphate-activated protein kinase; CAMKKβ, Ca++/calmodulin-dependent protein kinase-beta; DβH, dopamine-beta-hydroxylase; GR; type II glucocorticoid receptor; OVX, ovariectomy; pAMPK, phosphorylated AMPK; PP2A, protein phosphatase PP2A.

Article Snippet: pAMPK , Phosphorylated Thr 172 of human AMPK α1/2 , Santa Cruz Biotechnol.; rabbit; sc-33524; Lot no. J0915; AB_216714 , 0.4 ug/mL.

Techniques: Expressing