Structured Review

Millipore ampk specific inhibitor
Experimental design and animal groups. DHE: dihydroethidium; DMSO: dimethyl sulfoxide; <t>dorsomorphin:</t> <t>AMPK</t> inhibitor; Eze: Ezetimibe; IHC: immunohistochemistry; MCAO: middle cerebral artery occlusion; MDA: malonaldehyde; Scr siRNA: scramble siRNA; TTC: 2,3,5-triphenyltetrazolium chloride; WB: western blot.
Ampk Specific Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ampk specific inhibitor/product/Millipore
Average 99 stars, based on 1 article reviews
Price from $9.99 to $1999.99
ampk specific inhibitor - by Bioz Stars, 2021-07
99/100 stars

Images

1) Product Images from "Ezetimibe Attenuates Oxidative Stress and Neuroinflammation via the AMPK/Nrf2/TXNIP Pathway after MCAO in Rats"

Article Title: Ezetimibe Attenuates Oxidative Stress and Neuroinflammation via the AMPK/Nrf2/TXNIP Pathway after MCAO in Rats

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2020/4717258

Experimental design and animal groups. DHE: dihydroethidium; DMSO: dimethyl sulfoxide; dorsomorphin: AMPK inhibitor; Eze: Ezetimibe; IHC: immunohistochemistry; MCAO: middle cerebral artery occlusion; MDA: malonaldehyde; Scr siRNA: scramble siRNA; TTC: 2,3,5-triphenyltetrazolium chloride; WB: western blot.
Figure Legend Snippet: Experimental design and animal groups. DHE: dihydroethidium; DMSO: dimethyl sulfoxide; dorsomorphin: AMPK inhibitor; Eze: Ezetimibe; IHC: immunohistochemistry; MCAO: middle cerebral artery occlusion; MDA: malonaldehyde; Scr siRNA: scramble siRNA; TTC: 2,3,5-triphenyltetrazolium chloride; WB: western blot.

Techniques Used: Immunohistochemistry, Multiple Displacement Amplification, Western Blot

2) Product Images from "Metabolic intervention on lipid synthesis converging pathways abrogates prostate cancer growth"

Article Title: Metabolic intervention on lipid synthesis converging pathways abrogates prostate cancer growth

Journal: Oncogene

doi: 10.1038/onc.2012.523

Combined AMPK and FASN inhibition in PC cells is associated with an increased ME activity. ( a ) Induction of the SREBP-1 transcriptional activity following cC+ C75 co-treatment was assessed by measuring firefly Luc activity in LNCaP cells overexpressing SREBP-1 and FAS-Luc promoters, as compared with control. Values are expressed as relative luciferase unit (RLU) normalized relative to the β-galactosidase activity. ( b ) Variations in mRNA expression levels of SREBP-1 target genes G6PDH , ATP-citrate lyase , ME1 and malate dehydogenases ( Mdh )-1 and -2 were analyzed by RT-qPCR in LNCaP cells at 24 h following indicated treatments. ( c ) Induction of the ME1 transcriptional activity following cC+ C75 co-treatment was assessed by measuring Renilla luciferase (Luc) activity in LNCaP and C4-2 cells overexpressing ME1-Luc promoter, as compared with control. Values are expressed as relative luciferase unit (RLU) normalized relative to the β-galactosidase activity. ( d ) Induction of ME enzymatic activity in whole LNCaP cells was analyzed by spectrophotometry in the presence of NADPH substrate at 24 h following indicated treatments. ( e ) Rescue from the antiproliferative effect of cC+C75 treatment in LNCaP cells was conducted by genetic inhibition of ME1. ME mRNA and protein expression is indicated. ( f ) Quantification of the percentage of BrdU-incorporating cells transfected with siRNA against ME1, as compared with control.
Figure Legend Snippet: Combined AMPK and FASN inhibition in PC cells is associated with an increased ME activity. ( a ) Induction of the SREBP-1 transcriptional activity following cC+ C75 co-treatment was assessed by measuring firefly Luc activity in LNCaP cells overexpressing SREBP-1 and FAS-Luc promoters, as compared with control. Values are expressed as relative luciferase unit (RLU) normalized relative to the β-galactosidase activity. ( b ) Variations in mRNA expression levels of SREBP-1 target genes G6PDH , ATP-citrate lyase , ME1 and malate dehydogenases ( Mdh )-1 and -2 were analyzed by RT-qPCR in LNCaP cells at 24 h following indicated treatments. ( c ) Induction of the ME1 transcriptional activity following cC+ C75 co-treatment was assessed by measuring Renilla luciferase (Luc) activity in LNCaP and C4-2 cells overexpressing ME1-Luc promoter, as compared with control. Values are expressed as relative luciferase unit (RLU) normalized relative to the β-galactosidase activity. ( d ) Induction of ME enzymatic activity in whole LNCaP cells was analyzed by spectrophotometry in the presence of NADPH substrate at 24 h following indicated treatments. ( e ) Rescue from the antiproliferative effect of cC+C75 treatment in LNCaP cells was conducted by genetic inhibition of ME1. ME mRNA and protein expression is indicated. ( f ) Quantification of the percentage of BrdU-incorporating cells transfected with siRNA against ME1, as compared with control.

Techniques Used: Inhibition, Activity Assay, Luciferase, Expressing, Quantitative RT-PCR, Spectrophotometry, Transfection

Combined AMPK and FASN inhibition in PC cells is associated with an increased NOX activity and associated ROS production. ( a ) NADPH on NADP cellular ratio was measured in LNCaP cells by colorimetric assay at 48 h following indicated treatments. ( b ) NOX activity was measured by chemiluminescence in the presence of NADPH substrate and lucigenin at 24 h following indicated treatments. ( c ) NOX1 and -2 isoforms mRNA levels were analyzed by RT-qPCR at 24 h following indicated treatments. ( d , e ) FACS analysis of cellular ROS production measuring superoxide radicals (O 2 •− ) and H 2 O 2 with DHE ( d ) and DCFDA ( e ) fluoroprobes, respectively. ( f ) Co-treatment of cells with 2 μ M NOX inhibitor DPI abrogated cC+C75-induced ROS increase, as measured with DCFDA fluoroprobe by FACS analysis. ( g ) Rescue from NOX1 mRNA level induction following cC+C75 treatment was measured by RT-qPCR following LNCaP cell transfection with siRNA against ME1, as compared with control.
Figure Legend Snippet: Combined AMPK and FASN inhibition in PC cells is associated with an increased NOX activity and associated ROS production. ( a ) NADPH on NADP cellular ratio was measured in LNCaP cells by colorimetric assay at 48 h following indicated treatments. ( b ) NOX activity was measured by chemiluminescence in the presence of NADPH substrate and lucigenin at 24 h following indicated treatments. ( c ) NOX1 and -2 isoforms mRNA levels were analyzed by RT-qPCR at 24 h following indicated treatments. ( d , e ) FACS analysis of cellular ROS production measuring superoxide radicals (O 2 •− ) and H 2 O 2 with DHE ( d ) and DCFDA ( e ) fluoroprobes, respectively. ( f ) Co-treatment of cells with 2 μ M NOX inhibitor DPI abrogated cC+C75-induced ROS increase, as measured with DCFDA fluoroprobe by FACS analysis. ( g ) Rescue from NOX1 mRNA level induction following cC+C75 treatment was measured by RT-qPCR following LNCaP cell transfection with siRNA against ME1, as compared with control.

Techniques Used: Inhibition, Activity Assay, Colorimetric Assay, Quantitative RT-PCR, FACS, Transfection

Combined AMPK and FASN inhibition blocks PC cells' proliferation in vitro and induces caspase-dependent apoptosis. ( a ) Effects of cC and C75 inhibitors on LNCaP and C4-2 cell proliferation was assessed by cell counting at the indicated times. ( b – e ) Genetic inhibition of AMPK in PNT2, LNCaP and C4-2 cells was performed by nucleofaction with siRNA against hAMPKα1/2. Reduced AMPKα1/2 expression is shown by AMPKα1/2 immunoblotting ( b ) in LNCaP cells 24 h after transfection with control or siRNA against hAMPKα1/2. ( c ) Proliferation of sicont- or siAMPKα1/2—PNT2, LNCaP or C4-2 cells was measured by cell counting following treatment with control or C75-supplemented media at the indicated times. ( d ) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h following treatment with control or C75-supplemented media at the indicated times. Rescue from apoptosis induction was analyzed following pre-treatment of cells with 20 μ M qVD-oph caspase inhibitor. ( e ) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h after indicated siRNA treatments.
Figure Legend Snippet: Combined AMPK and FASN inhibition blocks PC cells' proliferation in vitro and induces caspase-dependent apoptosis. ( a ) Effects of cC and C75 inhibitors on LNCaP and C4-2 cell proliferation was assessed by cell counting at the indicated times. ( b – e ) Genetic inhibition of AMPK in PNT2, LNCaP and C4-2 cells was performed by nucleofaction with siRNA against hAMPKα1/2. Reduced AMPKα1/2 expression is shown by AMPKα1/2 immunoblotting ( b ) in LNCaP cells 24 h after transfection with control or siRNA against hAMPKα1/2. ( c ) Proliferation of sicont- or siAMPKα1/2—PNT2, LNCaP or C4-2 cells was measured by cell counting following treatment with control or C75-supplemented media at the indicated times. ( d ) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h following treatment with control or C75-supplemented media at the indicated times. Rescue from apoptosis induction was analyzed following pre-treatment of cells with 20 μ M qVD-oph caspase inhibitor. ( e ) Effect on cell viability was analyzed by FACS by measuring the percentage of annexin V-FITC+ cells at 16 h after indicated siRNA treatments.

Techniques Used: Inhibition, In Vitro, Cell Counting, Expressing, Transfection, FACS

Hypothetical mechanism underlying the proposed metabolic therapy. Inhibition of AMPK, in the context of inactive FASN, influences several metabolic pathways. First, AMPK inhibition activates ACC-mediated malonyl-CoA formation, the accumulation of which has apoptotic effects. Second, AMPK inhibition results in the activation of SREBP and of ME and G6PDH enzymes, resulting in the accumulation of NADPH. Moreover, FASN inhibition leads to further accumulation of NADPH, which is not used as a substrate for de novo lipids synthesis. As the NOX enzyme is also activated as a result of AMPK inhibition, excess of NADPH substrate will be converted into ROS. Altogether, this results in an increased apoptosis in cancer cells.
Figure Legend Snippet: Hypothetical mechanism underlying the proposed metabolic therapy. Inhibition of AMPK, in the context of inactive FASN, influences several metabolic pathways. First, AMPK inhibition activates ACC-mediated malonyl-CoA formation, the accumulation of which has apoptotic effects. Second, AMPK inhibition results in the activation of SREBP and of ME and G6PDH enzymes, resulting in the accumulation of NADPH. Moreover, FASN inhibition leads to further accumulation of NADPH, which is not used as a substrate for de novo lipids synthesis. As the NOX enzyme is also activated as a result of AMPK inhibition, excess of NADPH substrate will be converted into ROS. Altogether, this results in an increased apoptosis in cancer cells.

Techniques Used: Inhibition, Activation Assay

Combined AMPK and FASN inhibition reduces tumor growth in mice. ( a–c ) Tumor volume progression of subcutaneously implanted LNCaP ( a ), C4-2 ( b ) and PC-3 ( c ) cells in nude athymic mice was measured weekly following intraperitoneal injection with 10 mg/kg of either cC alone, C75 alone or both cC and C75 for 5 days a week. Values are expressed as the mean tumor volume (mm 3 )±s.e.m. ( n =5 for each group) from day 1 to day 14. ( d ) Ki67 and cleaved caspase-3 immunostaining of C4-2 tumor sections. ( e ) Quantification of cycling Ki67+ cells on C4-2 tumor sections. Six fields per section were analyzed for Ki67 immunostaining indicative of cell proliferation. Sections of tumors of all mice were analyzed. At least 300 cells were counted per tumor. ( f ) The lipidic composition was analyzed in total lipids extracted from C4-2 tumor tissues. Values are expressed as fold of control (mean±s.e.m.).
Figure Legend Snippet: Combined AMPK and FASN inhibition reduces tumor growth in mice. ( a–c ) Tumor volume progression of subcutaneously implanted LNCaP ( a ), C4-2 ( b ) and PC-3 ( c ) cells in nude athymic mice was measured weekly following intraperitoneal injection with 10 mg/kg of either cC alone, C75 alone or both cC and C75 for 5 days a week. Values are expressed as the mean tumor volume (mm 3 )±s.e.m. ( n =5 for each group) from day 1 to day 14. ( d ) Ki67 and cleaved caspase-3 immunostaining of C4-2 tumor sections. ( e ) Quantification of cycling Ki67+ cells on C4-2 tumor sections. Six fields per section were analyzed for Ki67 immunostaining indicative of cell proliferation. Sections of tumors of all mice were analyzed. At least 300 cells were counted per tumor. ( f ) The lipidic composition was analyzed in total lipids extracted from C4-2 tumor tissues. Values are expressed as fold of control (mean±s.e.m.).

Techniques Used: Inhibition, Mouse Assay, Injection, Immunostaining

Combined AMPK and FASN inhibition induces malonyl-CoA accumulation in PC cells. The AMPK and FASN inhibitors' concentration used in all in vitro experiments were as follows: cC 1 μg/ml, araA 0.5 m M and C75 7 μg/ml, unless otherwise indicated. ( a ) Immunoblot analysis of phospho (p)-AMPK and p-ACC in PNT2, LNCaP, C4-2 and PC-3 cell lines at 24 h following treatment with increasing concentration of AMPK activators AICAR or metformin (Metf.). ( b ) Immunoblot analysis of p-AMPK, total AMPK, p-ACC and tubulin in the LNCaP cell line at 24 h following treatment with either 1 μg/ml cC alone, 5–7 μg/ml C75 alone or with cC combined to increasing C75 concentrations. ( c ) Lipid synthesis through [ 14 C]palmitate incorporation into TAG was measured in C4-2 cells at 24 h following treatment with either cC, C75 or cC+C75 (left panel). Basal lipid synthesis levels were quantified in normal prostate PNT2 cells compared with C4-2 and PC-3 PC cells. ( d ) Cellular contents in malonyl-CoA and acetyl-CoA were measured in PNT2, LNCaP or C4-2 cells at 24 h following treatment with either cC, C75 or with combined cC+C75 treatment.
Figure Legend Snippet: Combined AMPK and FASN inhibition induces malonyl-CoA accumulation in PC cells. The AMPK and FASN inhibitors' concentration used in all in vitro experiments were as follows: cC 1 μg/ml, araA 0.5 m M and C75 7 μg/ml, unless otherwise indicated. ( a ) Immunoblot analysis of phospho (p)-AMPK and p-ACC in PNT2, LNCaP, C4-2 and PC-3 cell lines at 24 h following treatment with increasing concentration of AMPK activators AICAR or metformin (Metf.). ( b ) Immunoblot analysis of p-AMPK, total AMPK, p-ACC and tubulin in the LNCaP cell line at 24 h following treatment with either 1 μg/ml cC alone, 5–7 μg/ml C75 alone or with cC combined to increasing C75 concentrations. ( c ) Lipid synthesis through [ 14 C]palmitate incorporation into TAG was measured in C4-2 cells at 24 h following treatment with either cC, C75 or cC+C75 (left panel). Basal lipid synthesis levels were quantified in normal prostate PNT2 cells compared with C4-2 and PC-3 PC cells. ( d ) Cellular contents in malonyl-CoA and acetyl-CoA were measured in PNT2, LNCaP or C4-2 cells at 24 h following treatment with either cC, C75 or with combined cC+C75 treatment.

Techniques Used: Inhibition, Concentration Assay, In Vitro

Antiproliferative and proapoptotic effects of AMPK and FASN inhibition in PC cells are mediated by a NOX-dependent ROS overproduction. ( a ) The presence of oxidative stress as a consequence of ROS overproduction following cC+C75 treatment was analyzed by measuring the formation of thiobarbituric acid-reactive substrates (Tbars), indicative of lipid peroxidation, on LNCaP cellular extracts. Values are expressed as m M of malonaldehyde and normalized to protein content. ( b ) The mRNA expression levels of the antioxidant enzymes peroxidase (Perox)-1 and -2 and catalase, as measured by RT-qPCR, were compared following LNCaP treatment, as indicated. ( c – e ) Protective effects of antioxidants on ROS production ( c ), cell proliferation ( d ) and apoptosis ( e ) in cC+C75-treated LNCaP cells was conducted by 2-h pre-treatment with increasing amounts of the antioxidants ascorbic acid (AA) and Trolox, as indicated. ( c ) Rescue of ROS overproduction in cC+C75-treated LNCaP cells was assessed by FACS analysis by measuring H 2 O 2 content with DHE fluoroprobe following treatment with increasing doses of antioxidants. ( d ) Rescue of cC+C75-treated LNCaP cell proliferation was measured by XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2 H -tetrazolium-5-carboxanilide) assay at 48 h following co-treatment with increasing amounts of antioxidants. ( e ) Rescue of cC+C75-treated LNCaP cells from apoptosis induction was measured by FACS analysis of annexin V-FITC+ cells at 16 h following co-treatment with increasing amounts of antioxidants. ( f ) Rescue of cC+C75-treated LNCaP cells from apoptosis induction by 6-h co-treatment with 2 μ M NOX inhibitor DPI was measured by FACS analysis of annexin V-FITC+ cells.
Figure Legend Snippet: Antiproliferative and proapoptotic effects of AMPK and FASN inhibition in PC cells are mediated by a NOX-dependent ROS overproduction. ( a ) The presence of oxidative stress as a consequence of ROS overproduction following cC+C75 treatment was analyzed by measuring the formation of thiobarbituric acid-reactive substrates (Tbars), indicative of lipid peroxidation, on LNCaP cellular extracts. Values are expressed as m M of malonaldehyde and normalized to protein content. ( b ) The mRNA expression levels of the antioxidant enzymes peroxidase (Perox)-1 and -2 and catalase, as measured by RT-qPCR, were compared following LNCaP treatment, as indicated. ( c – e ) Protective effects of antioxidants on ROS production ( c ), cell proliferation ( d ) and apoptosis ( e ) in cC+C75-treated LNCaP cells was conducted by 2-h pre-treatment with increasing amounts of the antioxidants ascorbic acid (AA) and Trolox, as indicated. ( c ) Rescue of ROS overproduction in cC+C75-treated LNCaP cells was assessed by FACS analysis by measuring H 2 O 2 content with DHE fluoroprobe following treatment with increasing doses of antioxidants. ( d ) Rescue of cC+C75-treated LNCaP cell proliferation was measured by XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2 H -tetrazolium-5-carboxanilide) assay at 48 h following co-treatment with increasing amounts of antioxidants. ( e ) Rescue of cC+C75-treated LNCaP cells from apoptosis induction was measured by FACS analysis of annexin V-FITC+ cells at 16 h following co-treatment with increasing amounts of antioxidants. ( f ) Rescue of cC+C75-treated LNCaP cells from apoptosis induction by 6-h co-treatment with 2 μ M NOX inhibitor DPI was measured by FACS analysis of annexin V-FITC+ cells.

Techniques Used: Inhibition, Expressing, Quantitative RT-PCR, FACS

3) Product Images from "The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *"

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.486696

Med1-inducible liver cell proliferation is attenuated by AMPK inhibitor compound C. A and B , wild-type mice ( A ) and Ad-LacZ-injected mice ( B ) were given BrdUrd in drinking water for 3 days. C and D , mice were infected with Ad-Med1 by tail vein injection and given intraperitoneal injections of vehicle ( C ) or compound C ( D ) daily for 3 days with BrdUrd in drinking water. The livers of mice were processed for BrdUrd incorporation. E , quantification of BrdUrd-labeled nuclei shown in A–D . Nuclear labeling of hepatocytes indicates attenuation of Med-inducible hepatocyte proliferation by compound C.
Figure Legend Snippet: Med1-inducible liver cell proliferation is attenuated by AMPK inhibitor compound C. A and B , wild-type mice ( A ) and Ad-LacZ-injected mice ( B ) were given BrdUrd in drinking water for 3 days. C and D , mice were infected with Ad-Med1 by tail vein injection and given intraperitoneal injections of vehicle ( C ) or compound C ( D ) daily for 3 days with BrdUrd in drinking water. The livers of mice were processed for BrdUrd incorporation. E , quantification of BrdUrd-labeled nuclei shown in A–D . Nuclear labeling of hepatocytes indicates attenuation of Med-inducible hepatocyte proliferation by compound C.

Techniques Used: Mouse Assay, Injection, Infection, Labeling

Related Articles

Activity Assay:

Article Title: Enhanced Expression of WD Repeat-Containing Protein 35 via CaMKK/AMPK Activation in Bupivacaine-Treated Neuro2a Cells
Article Snippet: .. AMPK inhibitors, compound C and iodotubercidin, attenuate the bupivacaine-induced increase in AMPK and p38 MAPK activity in Neuro2a cells In order to explore the relationship between AMPK and p38 MAPK activation in bupivacaine-treated Neuro2a cells, cells were treated for 1 h with the AMPK inhibitor compound C (10 µM; Calbiochem, La Jolla, CA, USA) or iodotubercidin (1 µM; Abcam, Cambridge, MA, USA), followed by bupivacaine (2 mM) for 1 to 9 h. As shown in , compound C significantly attenuated the bupivacaine-induced increase in phosphorylation levels of AMPKα at time points from 3 to 9 h (P < 0.05 and P < 0.001). ..

Activation Assay:

Article Title: Enhanced Expression of WD Repeat-Containing Protein 35 via CaMKK/AMPK Activation in Bupivacaine-Treated Neuro2a Cells
Article Snippet: .. AMPK inhibitors, compound C and iodotubercidin, attenuate the bupivacaine-induced increase in AMPK and p38 MAPK activity in Neuro2a cells In order to explore the relationship between AMPK and p38 MAPK activation in bupivacaine-treated Neuro2a cells, cells were treated for 1 h with the AMPK inhibitor compound C (10 µM; Calbiochem, La Jolla, CA, USA) or iodotubercidin (1 µM; Abcam, Cambridge, MA, USA), followed by bupivacaine (2 mM) for 1 to 9 h. As shown in , compound C significantly attenuated the bupivacaine-induced increase in phosphorylation levels of AMPKα at time points from 3 to 9 h (P < 0.05 and P < 0.001). ..

Mouse Assay:

Article Title: Metabolic intervention on lipid synthesis converging pathways abrogates prostate cancer growth
Article Snippet: .. In each group, the mice were randomized according to their established tumor volume and given either the AMPK inhibitor cC at 10 mg/kg, the FASN inhibitor C75 at 10 mg/kg or both cC and C75 inhibitors (Sigma Aldrich) at 10 mg/kg, in 100 μl of a Labrafil-DMA-Tween-80 solution (89:10:1) (treated group) or vehicle alone (control group), by daily intraperitoneal injections 5 days a week. ..

In Vitro:

Article Title: Metformin promotes tau aggregation and exacerbates abnormal behavior in a mouse model of tauopathy
Article Snippet: Stock solutions of metformin, phenformin (Sigma), buformin (Santa Cruz) were prepared in water. .. Insulin (Humulin R® U-500, 100 UI/ml, Eli Lilly), heparin (12–15 kDa, 5000 UI/ml, Sirton Pharmaceuticals), rapamycin (LC labs; 25 mM in Ethanol), dorsomorphin (25 mM in DMSO), and okadaic acid (Sigma; 250 μM in DMSO) were used for experiments in vitro. ..

other:

Article Title: Quercetin-induced apoptosis ameliorates vascular smooth muscle cell senescence through AMP-activated protein kinase signaling pathway
Article Snippet: Compound C, an AMPK inhibitor, was provided by Calbiochem (La Jolla, CA, USA).

Acetylene Reduction Assay:

Article Title: Resistin promotes tumor metastasis by down-regulation of miR-519d through the AMPK/p38 signaling pathway in human chondrosarcoma cells
Article Snippet: ON-TARGETplus MMP2, p38, and control siRNAs were purchased from Dharmacon Research (Lafayette, CO). .. AMPK inhibitors (Ara A and compound C), p38 inhibitor (SB203580), MMP-2 inhibitor, and human MMP-2 ELISA kits were purchased from Calbiochem (San Diego, CA). .. Recombinant human resistin was purchased from PeproTech (Rocky Hill, NJ).

Enzyme-linked Immunosorbent Assay:

Article Title: Resistin promotes tumor metastasis by down-regulation of miR-519d through the AMPK/p38 signaling pathway in human chondrosarcoma cells
Article Snippet: ON-TARGETplus MMP2, p38, and control siRNAs were purchased from Dharmacon Research (Lafayette, CO). .. AMPK inhibitors (Ara A and compound C), p38 inhibitor (SB203580), MMP-2 inhibitor, and human MMP-2 ELISA kits were purchased from Calbiochem (San Diego, CA). .. Recombinant human resistin was purchased from PeproTech (Rocky Hill, NJ).

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    Millipore dorsomorphin
    BMP/Smad1/5/8 signaling pathway regulates Msx1 expression. A , immunofluorescence shows expressions of pp38 ( panel a ), pERK ( panel b ), and pJNK ( panel c ) in E13.5 molar tooth germ. B , in situ hybridization shows Msx1 expression in E12.5 molar germs after 12 h or 1 day in organ culture in the presence of DMSO ( panels a and d ), <t>dorsomorphin</t> ( panels b and e ), or SB203580 and U0126 ( panels c and f ). Panels a–c , whole mount in situ hybridization; panels d–f , tissue section in situ hybridization. C , a Western blot assay shows attenuation of Msx1 expression by dorsomorphin but not SB203580 and U0126 in BMP-induced primary dental mesenchymal cells from E13.5 embryos. GAPDH was used as internal control. Quantitative analysis is shown in the lower part. **, p
    Dorsomorphin, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dorsomorphin/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dorsomorphin - by Bioz Stars, 2021-07
    97/100 stars
      Buy from Supplier

    99
    Millipore ampk inhibitors
    AMP-activated protein kinase <t>(AMPK)</t> is involved in resistin-induced matrix metalloproteinase (MMP-2) expression and cell migration JJ012 and SW1353 cells were pre-treated with Ara A (0.5 mM) and compound C (10 μM) for 30 min or pre-transfected with control, AMPKα1, or AMPKα2 siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (A) Transwell assays, (B) enzyme-linked immunosorbent assay (ELISA), and (C) real-time quantitative polymerase chain reaction (RT-qPCR), respectively. Next, the JJ012 and SW1353 cells were pre-treated with SB203580 (10 μM) for 30 min or pre-transfected with control or <t>p38</t> siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (D) Transwell assays, (E) ELISA, and (F) RT-qPCR, respectively. (G) The JJ012 cells were incubated with resistin for the indicated time intervals, and the p-AMPK and p38 expression were examined by western blot. (H) The JJ012 cells were pre-treated for 30 min with Ara A and compound C, or SB203580 followed by stimulation with resistin. The p-p38 and p-AMPK expression were measured by western blot. The results are expressed as mean ± SEM. *, P
    Ampk Inhibitors, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ampk inhibitors/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ampk inhibitors - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    BMP/Smad1/5/8 signaling pathway regulates Msx1 expression. A , immunofluorescence shows expressions of pp38 ( panel a ), pERK ( panel b ), and pJNK ( panel c ) in E13.5 molar tooth germ. B , in situ hybridization shows Msx1 expression in E12.5 molar germs after 12 h or 1 day in organ culture in the presence of DMSO ( panels a and d ), dorsomorphin ( panels b and e ), or SB203580 and U0126 ( panels c and f ). Panels a–c , whole mount in situ hybridization; panels d–f , tissue section in situ hybridization. C , a Western blot assay shows attenuation of Msx1 expression by dorsomorphin but not SB203580 and U0126 in BMP-induced primary dental mesenchymal cells from E13.5 embryos. GAPDH was used as internal control. Quantitative analysis is shown in the lower part. **, p

    Journal: The Journal of Biological Chemistry

    Article Title: An Atypical Canonical Bone Morphogenetic Protein (BMP) Signaling Pathway Regulates Msh Homeobox 1 (Msx1) Expression during Odontogenesis *

    doi: 10.1074/jbc.M114.600064

    Figure Lengend Snippet: BMP/Smad1/5/8 signaling pathway regulates Msx1 expression. A , immunofluorescence shows expressions of pp38 ( panel a ), pERK ( panel b ), and pJNK ( panel c ) in E13.5 molar tooth germ. B , in situ hybridization shows Msx1 expression in E12.5 molar germs after 12 h or 1 day in organ culture in the presence of DMSO ( panels a and d ), dorsomorphin ( panels b and e ), or SB203580 and U0126 ( panels c and f ). Panels a–c , whole mount in situ hybridization; panels d–f , tissue section in situ hybridization. C , a Western blot assay shows attenuation of Msx1 expression by dorsomorphin but not SB203580 and U0126 in BMP-induced primary dental mesenchymal cells from E13.5 embryos. GAPDH was used as internal control. Quantitative analysis is shown in the lower part. **, p

    Article Snippet: For small molecule inhibition experiments, dorsomorphin (Sigma) or SB203580 (Cell Signaling) and U0126 (Cell Signaling) were added into the medium, respectively, at a final concentration of 20 μ m .

    Techniques: Expressing, Immunofluorescence, In Situ Hybridization, Organ Culture, Western Blot

    Structure of the ALK2-FKBP12 complex. A , secondary structure elements are labeled and shown as ribbons . Disordered parts of the L45 loop and A loop are indicated by a thin dashed line . Dorsomorphin is shown in gray stick representation, and Mg 2+ ion ( cyan ) and sulfate molecules ( purple ) are displayed as spheres. Inset box shows the specific interactions of dorsomorphin ( yellow sticks ) with the ATP pocket in ALK2. Hydrogen bonds ( blue dashed lines ) are formed with the hinge amide of His-286 and via a water molecule to Glu-248 (α C ). The planarity of the phenyl ring is restricted by the close packing of Val-214 and Gly-289 ( dashed gray lines). B , FKBP12 binding is dominated by insertion of the αGS2 helix into the central FK506-binding pocket of FKBP12. Here, the ALK5 residues Leu-195–Leu-196 are replaced by ALK2 Phe-198–Leu-199 resulting in a subtle shift in the complex interface. The hydrophobic contact surface in FKBP12 is colored yellow. C , | F o | − | F c | omit map contoured at 3σ ( green mesh ) for the bound dorsomorphin ligand ( yellow sticks ).

    Journal: The Journal of Biological Chemistry

    Article Title: Structure of the Bone Morphogenetic Protein Receptor ALK2 and Implications for Fibrodysplasia Ossificans Progressiva *

    doi: 10.1074/jbc.M112.365932

    Figure Lengend Snippet: Structure of the ALK2-FKBP12 complex. A , secondary structure elements are labeled and shown as ribbons . Disordered parts of the L45 loop and A loop are indicated by a thin dashed line . Dorsomorphin is shown in gray stick representation, and Mg 2+ ion ( cyan ) and sulfate molecules ( purple ) are displayed as spheres. Inset box shows the specific interactions of dorsomorphin ( yellow sticks ) with the ATP pocket in ALK2. Hydrogen bonds ( blue dashed lines ) are formed with the hinge amide of His-286 and via a water molecule to Glu-248 (α C ). The planarity of the phenyl ring is restricted by the close packing of Val-214 and Gly-289 ( dashed gray lines). B , FKBP12 binding is dominated by insertion of the αGS2 helix into the central FK506-binding pocket of FKBP12. Here, the ALK5 residues Leu-195–Leu-196 are replaced by ALK2 Phe-198–Leu-199 resulting in a subtle shift in the complex interface. The hydrophobic contact surface in FKBP12 is colored yellow. C , | F o | − | F c | omit map contoured at 3σ ( green mesh ) for the bound dorsomorphin ligand ( yellow sticks ).

    Article Snippet: The ALK2-FKBP12 complex was preincubated with 1 mm dorsomorphin (Calbiochem) at a protein concentration of 10 mg/ml and crystallized using a precipitant containing 30% PEG3350, 0.25 m ammonium sulfate, and 0.1 m BisTris, pH 6.0.

    Techniques: Labeling, Binding Assay

    Effects of dorsomorphin on Salmonella -induced changes in hepcidin expression and serum iron levels.

    Journal: The Journal of Clinical Investigation

    Article Title: Selective modulation of TLR4-activated inflammatory responses by altered iron homeostasis in mice

    doi: 10.1172/JCI39939

    Figure Lengend Snippet: Effects of dorsomorphin on Salmonella -induced changes in hepcidin expression and serum iron levels.

    Article Snippet: Treatment with dorsomorphin (12.5 μg/g BW i.p.; Calbiochem) or an equivalent volume of the vehicle DMSO was started at the same time as the streptomycin was administered and was continued twice daily for the duration of the experiment.

    Techniques: Expressing

    Effects of dorsomorphin treatment on Salmonella -induced intestinal inflammation in WT mice.

    Journal: The Journal of Clinical Investigation

    Article Title: Selective modulation of TLR4-activated inflammatory responses by altered iron homeostasis in mice

    doi: 10.1172/JCI39939

    Figure Lengend Snippet: Effects of dorsomorphin treatment on Salmonella -induced intestinal inflammation in WT mice.

    Article Snippet: Treatment with dorsomorphin (12.5 μg/g BW i.p.; Calbiochem) or an equivalent volume of the vehicle DMSO was started at the same time as the streptomycin was administered and was continued twice daily for the duration of the experiment.

    Techniques: Mouse Assay

    AMP-activated protein kinase (AMPK) is involved in resistin-induced matrix metalloproteinase (MMP-2) expression and cell migration JJ012 and SW1353 cells were pre-treated with Ara A (0.5 mM) and compound C (10 μM) for 30 min or pre-transfected with control, AMPKα1, or AMPKα2 siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (A) Transwell assays, (B) enzyme-linked immunosorbent assay (ELISA), and (C) real-time quantitative polymerase chain reaction (RT-qPCR), respectively. Next, the JJ012 and SW1353 cells were pre-treated with SB203580 (10 μM) for 30 min or pre-transfected with control or p38 siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (D) Transwell assays, (E) ELISA, and (F) RT-qPCR, respectively. (G) The JJ012 cells were incubated with resistin for the indicated time intervals, and the p-AMPK and p38 expression were examined by western blot. (H) The JJ012 cells were pre-treated for 30 min with Ara A and compound C, or SB203580 followed by stimulation with resistin. The p-p38 and p-AMPK expression were measured by western blot. The results are expressed as mean ± SEM. *, P

    Journal: Oncotarget

    Article Title: Resistin promotes tumor metastasis by down-regulation of miR-519d through the AMPK/p38 signaling pathway in human chondrosarcoma cells

    doi:

    Figure Lengend Snippet: AMP-activated protein kinase (AMPK) is involved in resistin-induced matrix metalloproteinase (MMP-2) expression and cell migration JJ012 and SW1353 cells were pre-treated with Ara A (0.5 mM) and compound C (10 μM) for 30 min or pre-transfected with control, AMPKα1, or AMPKα2 siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (A) Transwell assays, (B) enzyme-linked immunosorbent assay (ELISA), and (C) real-time quantitative polymerase chain reaction (RT-qPCR), respectively. Next, the JJ012 and SW1353 cells were pre-treated with SB203580 (10 μM) for 30 min or pre-transfected with control or p38 siRNA for 24 h, and subsequently stimulated with resistin (3 ng/ml) for 24 h. In vitro migration, MMP-2 protein expression (JJ012 cells), and MMP-2 mRNA expression were measured by (D) Transwell assays, (E) ELISA, and (F) RT-qPCR, respectively. (G) The JJ012 cells were incubated with resistin for the indicated time intervals, and the p-AMPK and p38 expression were examined by western blot. (H) The JJ012 cells were pre-treated for 30 min with Ara A and compound C, or SB203580 followed by stimulation with resistin. The p-p38 and p-AMPK expression were measured by western blot. The results are expressed as mean ± SEM. *, P

    Article Snippet: AMPK inhibitors (Ara A and compound C), p38 inhibitor (SB203580), MMP-2 inhibitor, and human MMP-2 ELISA kits were purchased from Calbiochem (San Diego, CA).

    Techniques: Expressing, Migration, Acetylene Reduction Assay, Transfection, In Vitro, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Incubation, Western Blot

    Resistin promotes cell migration and matrix metalloproteinase (MMP-2) expression by down-regulating microRNA (miR)-519d expression (A) The JJ012 and SW1353 cells were transfected with miR-519d mimic or inhibitor for 24 h, and cell migration ability was examined by Transwell assay. (B) The JJ012 cells were transfected with an miR-519d mimic or inhibitor for 24 h, and MMP-2 expression was examined by western blot (upper panel), and enzyme-linked immunosorbent assay (lower panel). (C) The JJ012 and SW1353 cells were incubated with resistin (0.3–3 ng/ml) for 24 h, and miR-519d expression was detected by real-time quantitative polymerase chain reaction. (D) Sequences of miR-519d and the potential miR-519d binding site at the MMP-2 3′ untranslated region (3′ UTR; WT-MMP-2 3′ UTR). Also shown are the nucleotides mutated in the MMP-2 3′UTR mutant (MUT-MMP2 3′ UTR). (E) The JJ012 and SW1353 cells were transfected with a wild-type or mutant MMP-2 3′ UTR luciferase plasmid for 24 h followed by stimulation with resistin (0.3-3 ng/ml) for 24 h, and the relative luciferase activity was measured. The JJ012 cells were pre-treated with AMPK and p38 inhibitors for 30 min or pre-transfected with specific siRNAs for 24 h followed by stimulation with resistin (3 ng/ml) for 24 h; and the (F, G) miR-519d expression and (H) MMP-2 3′ UTR activity were examined. The results are expressed as mean ± SEM. *, P

    Journal: Oncotarget

    Article Title: Resistin promotes tumor metastasis by down-regulation of miR-519d through the AMPK/p38 signaling pathway in human chondrosarcoma cells

    doi:

    Figure Lengend Snippet: Resistin promotes cell migration and matrix metalloproteinase (MMP-2) expression by down-regulating microRNA (miR)-519d expression (A) The JJ012 and SW1353 cells were transfected with miR-519d mimic or inhibitor for 24 h, and cell migration ability was examined by Transwell assay. (B) The JJ012 cells were transfected with an miR-519d mimic or inhibitor for 24 h, and MMP-2 expression was examined by western blot (upper panel), and enzyme-linked immunosorbent assay (lower panel). (C) The JJ012 and SW1353 cells were incubated with resistin (0.3–3 ng/ml) for 24 h, and miR-519d expression was detected by real-time quantitative polymerase chain reaction. (D) Sequences of miR-519d and the potential miR-519d binding site at the MMP-2 3′ untranslated region (3′ UTR; WT-MMP-2 3′ UTR). Also shown are the nucleotides mutated in the MMP-2 3′UTR mutant (MUT-MMP2 3′ UTR). (E) The JJ012 and SW1353 cells were transfected with a wild-type or mutant MMP-2 3′ UTR luciferase plasmid for 24 h followed by stimulation with resistin (0.3-3 ng/ml) for 24 h, and the relative luciferase activity was measured. The JJ012 cells were pre-treated with AMPK and p38 inhibitors for 30 min or pre-transfected with specific siRNAs for 24 h followed by stimulation with resistin (3 ng/ml) for 24 h; and the (F, G) miR-519d expression and (H) MMP-2 3′ UTR activity were examined. The results are expressed as mean ± SEM. *, P

    Article Snippet: AMPK inhibitors (Ara A and compound C), p38 inhibitor (SB203580), MMP-2 inhibitor, and human MMP-2 ELISA kits were purchased from Calbiochem (San Diego, CA).

    Techniques: Migration, Expressing, Transfection, Transwell Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Real-time Polymerase Chain Reaction, Binding Assay, Mutagenesis, Luciferase, Plasmid Preparation, Activity Assay