β actin antibodies  (Cell Signaling Technology Inc)

 
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    Name:
    β Actin 13E5 Rabbit mAb HRP Conjugate
    Description:
    This Cell Signaling Technology CST antibody is conjugated to the carbohydrate groups of horseradish peroxidase HRP via its amine groups The HRP conjugated antibody is expected to exhibit the same species cross reactivity as the unconjugated antibody β Actin 13E5 Rabbit mAb 4970
    Catalog Number:
    5125
    Price:
    None
    Category:
    Antibody Conjugates
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding residues near the amino terminus of human β-actin.
    Reactivity:
    Human Mouse Rat Monkey Bovine Pig
    Applications:
    Western Blot
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc β actin antibodies
    This Cell Signaling Technology CST antibody is conjugated to the carbohydrate groups of horseradish peroxidase HRP via its amine groups The HRP conjugated antibody is expected to exhibit the same species cross reactivity as the unconjugated antibody β Actin 13E5 Rabbit mAb 4970
    https://www.bioz.com/result/β actin antibodies/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    β actin antibodies - by Bioz Stars, 2021-03
    97/100 stars

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    Related Articles

    Western Blot:

    Article Title: Heterozygosity for Fibrinogen Results in Efficient Resolution of Kidney Ischemia Reperfusion Injury
    Article Snippet: Immunoblotting and Immunoprecipitation Kidney tissues were homogenized in RIPA buffer [50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% NP40, 1 mM PMSF, 1 mM NaF, 20 mM Na4 P2 O7 , 2 mM Na3 VO4 , 1X protease inhibitor cocktail (Roche Applied Science, Indianapolis, IN)] and equal protein (30 µg) was resolved by polyacrylamide gel electrophoresis. .. Proteins were transferred onto nitrocellulose membrane and western blotting was performed with rabbit polyclonal anti-fibrinogen (Dako), mouse monoclonal anti-pERK, anti-ERK2 (BD Biosciences San Diego, CA), anti-Cyclin D1, anti-pRb, anti-β -Actin (Cell Signaling Technology), anti-α-Tubulin (Sigma) and goat polyclonal HRP conjugated anti-mouse albumin (Abcam, Cambridge, MA). .. HRP conjugated secondary antibodies against mouse, rabbit and goat was purchased from Jackson Immunoresearch (West Grove, PA).

    Article Title: Resveratrol inhibits the phosphatidylinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in the human chronic myeloid leukemia K562 cell line
    Article Snippet: Resveratrol was further diluted in RPMI-1640 medium (Gibco, Big Cabin, OK, USA) plus 10% fetal bovine serum (FBS; Gibco) to the appropriate final concentrations. .. The primary polyclonal rabbit anti-human antibodies, anti-PI3K, -phosphorylated (p)-PI3K (Tyr458), -Akt, -p-Akt (Ser473), -mTOR, -p-mTOR (Ser2448), -p70S6K, -p-p70S6K (Thr389), -4EBP1, -p-4EBP1 (Ser65), -cyclin D1, -procaspase-3, -cleaved caspase-3 and-β-actin, were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA), and the secondary horseradish peroxidase (HRP)-labeled mouse anti-rabbit IgG polyclonal antibodies for western blot analysis were provided by Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. (Beijing, China). .. Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) were purchased from BD Biosciences (Palo Alto, CA, USA), LY294002 and SH-6 were provided by Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), and rapamycin (RAPA) was purchased from the North China Pharmaceutical Group Corporation (Shijiazhuang, China).

    Article Title: The expression and function of histamine H3 receptors in pancreatic beta cells
    Article Snippet: Samples were denatured at 75°C for 5 min. Aliquots containing 10 μg of protein were applied to 5–20% gradient polyacrylamide gels (Bio-Rad, Hercules, CA, USA) and separated by SDS-PAGE at constant voltage of 200 V for 30 min. After SDS-PAGE, the samples were transferred to a PVDF membrane by blotting at a constant current of 2.5 A for 3 min. .. The PVDF membrane was blocked by agitating in 5% non-fat dry milk solution for 1 h, then immunoblotted with rabbit anti-mouse H3 receptor antibody (0.4 μg·mL−1 in 5% BSA solution) or rabbit anti-β-actin antibody (Cell Signaling Technology, Danvers, MA, USA; 1:2000 dilution), and incubated at room temperature for 1 h. At the end of the incubation period, the membrane was washed then incubated with a HRP-labelled donkey anti-rabbit antibody (Thermo) as the secondary antibody (1:25 000 dilution in non-fat dry milk solution) at room temperature for 1 h. Immunoreactive bands were detected using an ECL Western Blotting Analysis System (PerkinElmer, Waltham, MA, USA), and bands were recorded using the Ez-capture MG chemiluminescence imaging system (Atto, Tokyo, Japan). .. Pancreata were quickly removed from 12-week-old WT ( n = 5), H3 KO ( n = 3) and HdcKO ( n = 3) mice.

    Incubation:

    Article Title: Dispatched conformational dynamics couples transmembrane Na+ flux to release of lipid-modified Hedgehog signal
    Article Snippet: Western blotting was performed using Criterion TGX stain-Free precast gels (Bio-rad) and Immobilon-P PVDF membrane (Millipore). .. PVDF membranes were immunoblotted overnight at 4°C with anti-HA rabbit polyclonal (1:1000, Cell signaling) or anti-β-actin rabbit polyclonal (1:1000, Cell signaling) antibodies followed by a 1-hour incubation with the corresponding HRP-conjugated secondary antibodies (1:2000, Promega) at room temperature. .. The blots were developed using SuperSignal West Pico PLUS substrate (ThermoFisher).

    Article Title: The expression and function of histamine H3 receptors in pancreatic beta cells
    Article Snippet: Samples were denatured at 75°C for 5 min. Aliquots containing 10 μg of protein were applied to 5–20% gradient polyacrylamide gels (Bio-Rad, Hercules, CA, USA) and separated by SDS-PAGE at constant voltage of 200 V for 30 min. After SDS-PAGE, the samples were transferred to a PVDF membrane by blotting at a constant current of 2.5 A for 3 min. .. The PVDF membrane was blocked by agitating in 5% non-fat dry milk solution for 1 h, then immunoblotted with rabbit anti-mouse H3 receptor antibody (0.4 μg·mL−1 in 5% BSA solution) or rabbit anti-β-actin antibody (Cell Signaling Technology, Danvers, MA, USA; 1:2000 dilution), and incubated at room temperature for 1 h. At the end of the incubation period, the membrane was washed then incubated with a HRP-labelled donkey anti-rabbit antibody (Thermo) as the secondary antibody (1:25 000 dilution in non-fat dry milk solution) at room temperature for 1 h. Immunoreactive bands were detected using an ECL Western Blotting Analysis System (PerkinElmer, Waltham, MA, USA), and bands were recorded using the Ez-capture MG chemiluminescence imaging system (Atto, Tokyo, Japan). .. Pancreata were quickly removed from 12-week-old WT ( n = 5), H3 KO ( n = 3) and HdcKO ( n = 3) mice.

    other:

    Article Title: Pyroptosis is a critical inflammatory pathway in the placenta from early onset preeclampsia and in human trophoblasts exposed to hypoxia and endoplasmic reticulum stressors
    Article Snippet: Antibodies and reagents The following antibodies were used: rabbit anti-TXNIP (Cell Signaling, #14715), rabbit anti- PERK (Cell Signaling, #5683), rabbit anti-IL-18 (Abcam, ab68435), rabbit anti-NLRP3 (Abcam, ab214185), rabbit anti-IL-1β (Santa Cruz, sc-7884), rabbit anti-caspase-1 (Cell Signaling, #2225), mouse anti-Gasdermin D (Abcam, ab57785), mouse anti-β-actin (Cell Signaling), rabbit anti-BiP (Abcam, ab21685), rabbit anti-IRE1a (Abcam, ab48187), rabbit anti-phospho-MLKL (Ser358) (Cell Signaling, D6H3V), rat anti-MLKL (Millipore, MABC604), mouse anti-ASC (B-3) (Santa Cruz, sc-514414), rabbit anti-cleaved gasdermin D (Asp275) (Cell Signaling, #36425), goat anti-rabbit or mouse HRP-conjugated IgG (Cell Signaling), Alexa Fluor 488 donkey anti-rabbit or mouse IgG (Molecular Probes, A-21206), and Alexa Fluor 594 donkey anti-mouse IgG (Molecular Probes, A-21203).

    Article Title: Paneth cells as a site of origin for intestinal inflammation
    Article Snippet: Cell Signaling Technology: anti-β-actin (4970; 13E5) anti-GAPDH (2118; 14C10), anti-eIF2α (9722), anti-phospho-eIF2α (3597; 119A11), anti-PERK (3192; C33E10), anti-phospho-PERK (3179; 16F8), anti-JNK (9252), anti-phospho-JNK (4668; 81E11), anti-ATG5 (8540; D1G9), anti-Beclin-1 (3495; D40C5), anti-ATG7 (8558; D12B11), anti-CHOP (5554; D46F1), anti-ATG12 (4180; D88H11), anti-p62 (5114), anti-IKK1 (2682), anti-IKK2 (2370; 2C8), anti-phospho-IKK1/2 (2697; 16A6), anti-IRE1α (3294; 14C10), anti-phospho-NFκB p65 (33033; 93H1), anti-NFκB p65 (4764; C22B4) and anti-rabbit/mouse HRP antibodies (7074, 7076).

    Imaging:

    Article Title: The expression and function of histamine H3 receptors in pancreatic beta cells
    Article Snippet: Samples were denatured at 75°C for 5 min. Aliquots containing 10 μg of protein were applied to 5–20% gradient polyacrylamide gels (Bio-Rad, Hercules, CA, USA) and separated by SDS-PAGE at constant voltage of 200 V for 30 min. After SDS-PAGE, the samples were transferred to a PVDF membrane by blotting at a constant current of 2.5 A for 3 min. .. The PVDF membrane was blocked by agitating in 5% non-fat dry milk solution for 1 h, then immunoblotted with rabbit anti-mouse H3 receptor antibody (0.4 μg·mL−1 in 5% BSA solution) or rabbit anti-β-actin antibody (Cell Signaling Technology, Danvers, MA, USA; 1:2000 dilution), and incubated at room temperature for 1 h. At the end of the incubation period, the membrane was washed then incubated with a HRP-labelled donkey anti-rabbit antibody (Thermo) as the secondary antibody (1:25 000 dilution in non-fat dry milk solution) at room temperature for 1 h. Immunoreactive bands were detected using an ECL Western Blotting Analysis System (PerkinElmer, Waltham, MA, USA), and bands were recorded using the Ez-capture MG chemiluminescence imaging system (Atto, Tokyo, Japan). .. Pancreata were quickly removed from 12-week-old WT ( n = 5), H3 KO ( n = 3) and HdcKO ( n = 3) mice.

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  • 86
    Cell Signaling Technology Inc rabbit anti β actin
    Genetic ablation of Nrf2 reduces angiogenic sprouting and vascular density. ( A ) Blood vessels were visualized by PECAM-1 staining of retinas at P5. Decreased vascular density ( B ), branch points ( C ), sprouts ( D ), and filopodia ( E ) were observed in Nrf2 − / − retina ( n = 6). ( F and G ) Isolectin B4 (green) and BrdU labeling (red) of retinas at P5 demonstrate reduced EC proliferation in Nrf2 − / − ( n = 4). (Scale bar, 50 µm.) ( H ) NG2 + pericytes in capillaries or SMA + vascular smooth muscle cells in arterioles were indistinguishable between WT ( Nrf2 +/+ ) and Nrf2 − / − retinas at P5. (Scale bar, 100 µm.) ( I ) Nrf2 is expressed in developing blood vessels, and strong expression was observed in the sprouting vascular front. Arrowheads indicate Nrf2 expression in tip cells and stalk cells (A, artery; V, vein). ( J ) Nrf2 expression was increased in the ganglion cell layer (GCL) of retina at P5 compared with P0. Nrf2 was highly expressed in the vessels and surrounding area (arrowheads). Nrf2 localization in the nucleus was apparent (arrows). HV, hyaloid vessels; INL, inner nuclear layer. ( K ) Immunoblot analysis of Nrf2 in the retinas of WT mice at P0 and P5. <t>β-Actin</t> was used as a loading control ( n = 3). ( L ) Quantitative RT-PCR analysis of Nrf2 and Nrf2 target genes in the retinas of WT mice at P0 and P5 ( n = 5). Data are presented as mean ± SEM (** P
    Rabbit Anti β Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti β actin/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    86
    Cell Signaling Technology Inc ãŽâ² actin
    Mediator complex subunit 1 (Med1) is regulated by microRNA (miR)-96/182/183 in mice. A : expression of Med1 by real-time PCR in mouse primary hepatocytes transfected with 50 nM of Med1 or scramble siRNA; n = 3 B : Western blot of Med1 and <t>β-actin</t> (loading control) from protein samples of primary hepatocytes transfected with Med1 or scramble siRNA. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. A.U., arbitrary units. C : hepatic Med1 mRNA levels in db/db mice treated with vehicle or 2% colesevelam (Col)-supplemented diet and PBS or locked-nucleic acid (LNA)-182-5p; real-time PCR; n = 8–9. D : hepatic Med1 and β-actin (loading control) protein levels from 3 representative liver protein samples for Chow + PBS, Col + PBS, and Col + LNA-182. Western blotting. Ratio of Med1/β-actin densitometry is indicated between the blots. E : densitometry quantification of Med1 protein levels normalized to β-actin; n = 8–9. F : expression of miR-96-5p, miR-182-5p, and miR-183-5p in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, miR-183-5p mimics individually or in combination; real-time PCR; n = 3. G : Med1 mRNA levels in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 3. H : protein levels of Med1 and β-actin (loading control) from protein samples of primary hepatocytes transfected with mock or 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. For comparisons between 2 groups, Students’ t- tests were used, and for more than 2 groups, Kruskal-Wallis one-way ANOVA with Dunn’s posttest was used (α = 0.05) or one-way ANOVA with Bonferonni’s posttest for comparison to mock group only was used (α = 0.05).
    ãŽâ² Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc rabbit monoclonal antibodies against β actin
    HG treatment suppresses the expression of PRKAA1 and upregulates levels of miR-137 and IL-6. HTR-8/SVneo trophoblastic cells were treated with HG (25 mmol/l) medium for 24 h, with a control group cultured in normal medium (5 mmol/l). Expression levels of miR-137 and PRKAA1 were detected using reverse transcription-quantitative polymerase chain reaction analysis, with hsa-u6 and <t>β-actin</t> as internal controls. Protein levels of PRKAA1 (~62 kDa) and IL-6 were measured using western blot analysis and ELISA, respectively, and the relative density of PRKAA1 was determined using Image Pro Plus 6.0 software. (A) HG intervention suppressed the expression of PRKAA1 in HTR-8/SVneo cells. (B) miR-137 expression levels were elevated in HG-exposed HTR-8/SVneo cells. (C) Protein level of PRKAA1 was decreased in HG-exposed HTR-8/SVneo cells. (D) Results of western blot analysis. (E) HG treatment promoted the secretion of IL-6 in HTR-8/SVneo cells. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P
    Rabbit Monoclonal Antibodies Against β Actin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Genetic ablation of Nrf2 reduces angiogenic sprouting and vascular density. ( A ) Blood vessels were visualized by PECAM-1 staining of retinas at P5. Decreased vascular density ( B ), branch points ( C ), sprouts ( D ), and filopodia ( E ) were observed in Nrf2 − / − retina ( n = 6). ( F and G ) Isolectin B4 (green) and BrdU labeling (red) of retinas at P5 demonstrate reduced EC proliferation in Nrf2 − / − ( n = 4). (Scale bar, 50 µm.) ( H ) NG2 + pericytes in capillaries or SMA + vascular smooth muscle cells in arterioles were indistinguishable between WT ( Nrf2 +/+ ) and Nrf2 − / − retinas at P5. (Scale bar, 100 µm.) ( I ) Nrf2 is expressed in developing blood vessels, and strong expression was observed in the sprouting vascular front. Arrowheads indicate Nrf2 expression in tip cells and stalk cells (A, artery; V, vein). ( J ) Nrf2 expression was increased in the ganglion cell layer (GCL) of retina at P5 compared with P0. Nrf2 was highly expressed in the vessels and surrounding area (arrowheads). Nrf2 localization in the nucleus was apparent (arrows). HV, hyaloid vessels; INL, inner nuclear layer. ( K ) Immunoblot analysis of Nrf2 in the retinas of WT mice at P0 and P5. β-Actin was used as a loading control ( n = 3). ( L ) Quantitative RT-PCR analysis of Nrf2 and Nrf2 target genes in the retinas of WT mice at P0 and P5 ( n = 5). Data are presented as mean ± SEM (** P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Nrf2 acts cell-autonomously in endothelium to regulate tip cell formation and vascular branching

    doi: 10.1073/pnas.1309276110

    Figure Lengend Snippet: Genetic ablation of Nrf2 reduces angiogenic sprouting and vascular density. ( A ) Blood vessels were visualized by PECAM-1 staining of retinas at P5. Decreased vascular density ( B ), branch points ( C ), sprouts ( D ), and filopodia ( E ) were observed in Nrf2 − / − retina ( n = 6). ( F and G ) Isolectin B4 (green) and BrdU labeling (red) of retinas at P5 demonstrate reduced EC proliferation in Nrf2 − / − ( n = 4). (Scale bar, 50 µm.) ( H ) NG2 + pericytes in capillaries or SMA + vascular smooth muscle cells in arterioles were indistinguishable between WT ( Nrf2 +/+ ) and Nrf2 − / − retinas at P5. (Scale bar, 100 µm.) ( I ) Nrf2 is expressed in developing blood vessels, and strong expression was observed in the sprouting vascular front. Arrowheads indicate Nrf2 expression in tip cells and stalk cells (A, artery; V, vein). ( J ) Nrf2 expression was increased in the ganglion cell layer (GCL) of retina at P5 compared with P0. Nrf2 was highly expressed in the vessels and surrounding area (arrowheads). Nrf2 localization in the nucleus was apparent (arrows). HV, hyaloid vessels; INL, inner nuclear layer. ( K ) Immunoblot analysis of Nrf2 in the retinas of WT mice at P0 and P5. β-Actin was used as a loading control ( n = 3). ( L ) Quantitative RT-PCR analysis of Nrf2 and Nrf2 target genes in the retinas of WT mice at P0 and P5 ( n = 5). Data are presented as mean ± SEM (** P

    Article Snippet: The following antibodies were used: rabbit anti-Nrf2, mouse anti-GAPDH (Abcam), rabbit anti–β-actin, rabbit anti-NICD, rabbit anti–phospho-Akt, mouse anti-Akt1, mouse anti–phospho-p44/42 MAPK (Erk1/2), rabbit anti-p44/42 MAPK (Erk1/2), and rabbit anti-Dll4 (Cell Signaling Technology).

    Techniques: Staining, Labeling, Expressing, Mouse Assay, Quantitative RT-PCR

    Western blot analysis of poly (adenosine diphosphate-ribose) polymerase in A549, PC3 and H1299 cell lines. β-actin was used as loading control. The treatment was conducted for 24 h. Vertical and horizontal lanes indicate separate blots. PQE, Pelargonium quercetorum extract; PARP, poly (adenosine diphosphate-ribose) polymerase.

    Journal: Oncology Letters

    Article Title: Pelargonium quercetorum Agnew induces apoptosis without PARP or cytokeratin 18 cleavage in non-small cell lung cancer cell lines

    doi: 10.3892/ol.2016.4779

    Figure Lengend Snippet: Western blot analysis of poly (adenosine diphosphate-ribose) polymerase in A549, PC3 and H1299 cell lines. β-actin was used as loading control. The treatment was conducted for 24 h. Vertical and horizontal lanes indicate separate blots. PQE, Pelargonium quercetorum extract; PARP, poly (adenosine diphosphate-ribose) polymerase.

    Article Snippet: Western blotting was performed using rabbit anti-β-actin and anti-PARP monoclonal antibodies at 1:1,000 dilution (catalog nos., 4970 and 9532, respectively; Cell Signaling Technology, Inc., Danvers, MA, USA) in 5% (w/v) bovine serum albumin (Amresco, LLC, Solon, OH, USA).

    Techniques: Western Blot

    Mediator complex subunit 1 (Med1) is regulated by microRNA (miR)-96/182/183 in mice. A : expression of Med1 by real-time PCR in mouse primary hepatocytes transfected with 50 nM of Med1 or scramble siRNA; n = 3 B : Western blot of Med1 and β-actin (loading control) from protein samples of primary hepatocytes transfected with Med1 or scramble siRNA. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. A.U., arbitrary units. C : hepatic Med1 mRNA levels in db/db mice treated with vehicle or 2% colesevelam (Col)-supplemented diet and PBS or locked-nucleic acid (LNA)-182-5p; real-time PCR; n = 8–9. D : hepatic Med1 and β-actin (loading control) protein levels from 3 representative liver protein samples for Chow + PBS, Col + PBS, and Col + LNA-182. Western blotting. Ratio of Med1/β-actin densitometry is indicated between the blots. E : densitometry quantification of Med1 protein levels normalized to β-actin; n = 8–9. F : expression of miR-96-5p, miR-182-5p, and miR-183-5p in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, miR-183-5p mimics individually or in combination; real-time PCR; n = 3. G : Med1 mRNA levels in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 3. H : protein levels of Med1 and β-actin (loading control) from protein samples of primary hepatocytes transfected with mock or 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. For comparisons between 2 groups, Students’ t- tests were used, and for more than 2 groups, Kruskal-Wallis one-way ANOVA with Dunn’s posttest was used (α = 0.05) or one-way ANOVA with Bonferonni’s posttest for comparison to mock group only was used (α = 0.05).

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Liver and Biliary Tract Physiology/Pathophysiology: Intestinal bile acid sequestration improves glucose control by stimulating hepatic miR-182-5p in type 2 diabetes

    doi: 10.1152/ajpgi.00238.2018

    Figure Lengend Snippet: Mediator complex subunit 1 (Med1) is regulated by microRNA (miR)-96/182/183 in mice. A : expression of Med1 by real-time PCR in mouse primary hepatocytes transfected with 50 nM of Med1 or scramble siRNA; n = 3 B : Western blot of Med1 and β-actin (loading control) from protein samples of primary hepatocytes transfected with Med1 or scramble siRNA. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. A.U., arbitrary units. C : hepatic Med1 mRNA levels in db/db mice treated with vehicle or 2% colesevelam (Col)-supplemented diet and PBS or locked-nucleic acid (LNA)-182-5p; real-time PCR; n = 8–9. D : hepatic Med1 and β-actin (loading control) protein levels from 3 representative liver protein samples for Chow + PBS, Col + PBS, and Col + LNA-182. Western blotting. Ratio of Med1/β-actin densitometry is indicated between the blots. E : densitometry quantification of Med1 protein levels normalized to β-actin; n = 8–9. F : expression of miR-96-5p, miR-182-5p, and miR-183-5p in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, miR-183-5p mimics individually or in combination; real-time PCR; n = 3. G : Med1 mRNA levels in mouse primary hepatocytes transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 3. H : protein levels of Med1 and β-actin (loading control) from protein samples of primary hepatocytes transfected with mock or 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 3. For comparisons between 2 groups, Students’ t- tests were used, and for more than 2 groups, Kruskal-Wallis one-way ANOVA with Dunn’s posttest was used (α = 0.05) or one-way ANOVA with Bonferonni’s posttest for comparison to mock group only was used (α = 0.05).

    Article Snippet: Rabbit anti-mouse antibodies against MED1 (lot 2G073735; Abnova PAB8661; polyclonal, 1:1,000 dilution in Odyssey blocking buffer) and β-actin (lot 14; Cell Signaling 13E5; monoclonal, 1:5,000 dilution in Odyssey blocking buffer) were used for Western blotting and detected using the LICOR system.

    Techniques: Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot

    Colesevelam (Col) alters hepatic lipid metabolism but has modest effects on incretins. A : levels of glucagon like peptide-1 (GLP-1) in portal blood from overnight-fasted animals; n = 24. B : levels of gastric inhibitory peptide (GIP) in portal blood from overnight-fasted animals; n = 24. C : plasma GIP levels after intraperitena glucose challenge; n = 12. D : percentage of new islets by 5-bromo-2'-deoxyuridine labeling; n = 12. E : homeostatic model assessment (HOMA) method for assessing insulin resistance from basal (fasting) glucose and insulin concentrations; n = 22–24 F : homeostatic model assessment (HOMA) method for assessing insulin β-cell function from basal (fasting) glucose and C-peptide concentrations; n = 22–23. G : percent de novo cholesterol synthesis over 4 wk; n = 6. H : de novo lipogenesis, percent new triglyceride-palmitate after 4 wk of labeling; n = 6. For comparisons between 2 groups, Mann-Whitney nonparametric tests were used, and for repeated measures across time, two-way ANOVAs with Bonferonni’s posttests were used.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Liver and Biliary Tract Physiology/Pathophysiology: Intestinal bile acid sequestration improves glucose control by stimulating hepatic miR-182-5p in type 2 diabetes

    doi: 10.1152/ajpgi.00238.2018

    Figure Lengend Snippet: Colesevelam (Col) alters hepatic lipid metabolism but has modest effects on incretins. A : levels of glucagon like peptide-1 (GLP-1) in portal blood from overnight-fasted animals; n = 24. B : levels of gastric inhibitory peptide (GIP) in portal blood from overnight-fasted animals; n = 24. C : plasma GIP levels after intraperitena glucose challenge; n = 12. D : percentage of new islets by 5-bromo-2'-deoxyuridine labeling; n = 12. E : homeostatic model assessment (HOMA) method for assessing insulin resistance from basal (fasting) glucose and insulin concentrations; n = 22–24 F : homeostatic model assessment (HOMA) method for assessing insulin β-cell function from basal (fasting) glucose and C-peptide concentrations; n = 22–23. G : percent de novo cholesterol synthesis over 4 wk; n = 6. H : de novo lipogenesis, percent new triglyceride-palmitate after 4 wk of labeling; n = 6. For comparisons between 2 groups, Mann-Whitney nonparametric tests were used, and for repeated measures across time, two-way ANOVAs with Bonferonni’s posttests were used.

    Article Snippet: Rabbit anti-mouse antibodies against MED1 (lot 2G073735; Abnova PAB8661; polyclonal, 1:1,000 dilution in Odyssey blocking buffer) and β-actin (lot 14; Cell Signaling 13E5; monoclonal, 1:5,000 dilution in Odyssey blocking buffer) were used for Western blotting and detected using the LICOR system.

    Techniques: Labeling, Cell Function Assay, MANN-WHITNEY

    Mediator complex subunit 1 (MED1) is a direct target of microRNA (miR)-182-5p, miR-183-5p, and miR-96-5p in humans. A : schematic of human MED1 mRNA and the 3 putative miR-182/183/96 target sites in the 3′-untranslated region (3′-UTR). B : predicted MED1 3′-UTR target sites. Bases highlighted in dark gray represent mutations for gene reporter (luciferase) assays. C : normalized luciferase activity in HEK293 cells after dual transfection of miR-96/182/183 mimics and gene (luciferase) reporters harboring putative target sites for miR-96-5p, miR-182-5p, and/or miR-183-5p; n = 4. D : miR-96-5p, miR-182-5p, and miR-183-5p levels in Huh7 cells transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 6. E : MED1 mRNA levels in Huh7 cells transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 6. F : miR-96-5p, miR-182-5p, and miR-183-5p levels in Huh7 cells transfected with mock or 50 nM locked-nucleic acids (LNAs) against miR-182-5p and/or miR-183-5p; real-time PCR; n = 6. G : MED1 mRNA levels in Huh7 cells transfected mock or 50 nM LNAs against miR-182-5p and/or miR-183-5p; real-time PCR; n = 6. H : protein levels of Med1 and β-actin (loading control) from protein lysates of Huh7 cells transfected with mock or 50 nM miR-96, miR-182, and miR-183 LNAs. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 6. A.U. arbitrary units. For comparisons between more than 2 groups to mock only, one-way ANOVA with Bonferonni’s posttest was used, α = 0.05. For comparisons between 2 groups, Mann-Whitney nonparametric tests were used.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Liver and Biliary Tract Physiology/Pathophysiology: Intestinal bile acid sequestration improves glucose control by stimulating hepatic miR-182-5p in type 2 diabetes

    doi: 10.1152/ajpgi.00238.2018

    Figure Lengend Snippet: Mediator complex subunit 1 (MED1) is a direct target of microRNA (miR)-182-5p, miR-183-5p, and miR-96-5p in humans. A : schematic of human MED1 mRNA and the 3 putative miR-182/183/96 target sites in the 3′-untranslated region (3′-UTR). B : predicted MED1 3′-UTR target sites. Bases highlighted in dark gray represent mutations for gene reporter (luciferase) assays. C : normalized luciferase activity in HEK293 cells after dual transfection of miR-96/182/183 mimics and gene (luciferase) reporters harboring putative target sites for miR-96-5p, miR-182-5p, and/or miR-183-5p; n = 4. D : miR-96-5p, miR-182-5p, and miR-183-5p levels in Huh7 cells transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 6. E : MED1 mRNA levels in Huh7 cells transfected with 50 nM miR-96-5p, miR-182-5p, and miR-183-5p mimics individually or in combination; real-time PCR; n = 6. F : miR-96-5p, miR-182-5p, and miR-183-5p levels in Huh7 cells transfected with mock or 50 nM locked-nucleic acids (LNAs) against miR-182-5p and/or miR-183-5p; real-time PCR; n = 6. G : MED1 mRNA levels in Huh7 cells transfected mock or 50 nM LNAs against miR-182-5p and/or miR-183-5p; real-time PCR; n = 6. H : protein levels of Med1 and β-actin (loading control) from protein lysates of Huh7 cells transfected with mock or 50 nM miR-96, miR-182, and miR-183 LNAs. Ratio of Med1/β-actin densitometry is indicated between the blots and graphed on the right ; n = 6. A.U. arbitrary units. For comparisons between more than 2 groups to mock only, one-way ANOVA with Bonferonni’s posttest was used, α = 0.05. For comparisons between 2 groups, Mann-Whitney nonparametric tests were used.

    Article Snippet: Rabbit anti-mouse antibodies against MED1 (lot 2G073735; Abnova PAB8661; polyclonal, 1:1,000 dilution in Odyssey blocking buffer) and β-actin (lot 14; Cell Signaling 13E5; monoclonal, 1:5,000 dilution in Odyssey blocking buffer) were used for Western blotting and detected using the LICOR system.

    Techniques: Luciferase, Activity Assay, Transfection, Real-time Polymerase Chain Reaction, MANN-WHITNEY

    Colesevelam (Col) stimulates the microRNA miR-96/182/183 cluster in livers of db/db mice and is inhibited with locked-nucleic acid (LNA)-182 treatment. A : schematic of animal study. Twelve-week-old db/db mice treated with vehicle or 2% colesevelam-supplemented diet for 9 wk. GTT, glucose tolerance test; ITT, insulin tolerance test. B : liver expression of miR-96/182/183 cluster miRNAs; n = 8–9. C : liver gene (mRNA) expression changes for SREBP2 target genes Hmgcr , Ldlr , Sqle , and Srebf2 ; n = 8–9. Actb, β-actin. D : white adipose tissue (WAT) miRNA expression of miR-182/183/96 cluster; n = 8–9. For comparisons among 3 groups, Kruskal-Wallis one-way ANOVA with Dunn’s posttest was used; α = 0.05.

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Liver and Biliary Tract Physiology/Pathophysiology: Intestinal bile acid sequestration improves glucose control by stimulating hepatic miR-182-5p in type 2 diabetes

    doi: 10.1152/ajpgi.00238.2018

    Figure Lengend Snippet: Colesevelam (Col) stimulates the microRNA miR-96/182/183 cluster in livers of db/db mice and is inhibited with locked-nucleic acid (LNA)-182 treatment. A : schematic of animal study. Twelve-week-old db/db mice treated with vehicle or 2% colesevelam-supplemented diet for 9 wk. GTT, glucose tolerance test; ITT, insulin tolerance test. B : liver expression of miR-96/182/183 cluster miRNAs; n = 8–9. C : liver gene (mRNA) expression changes for SREBP2 target genes Hmgcr , Ldlr , Sqle , and Srebf2 ; n = 8–9. Actb, β-actin. D : white adipose tissue (WAT) miRNA expression of miR-182/183/96 cluster; n = 8–9. For comparisons among 3 groups, Kruskal-Wallis one-way ANOVA with Dunn’s posttest was used; α = 0.05.

    Article Snippet: Rabbit anti-mouse antibodies against MED1 (lot 2G073735; Abnova PAB8661; polyclonal, 1:1,000 dilution in Odyssey blocking buffer) and β-actin (lot 14; Cell Signaling 13E5; monoclonal, 1:5,000 dilution in Odyssey blocking buffer) were used for Western blotting and detected using the LICOR system.

    Techniques: Mouse Assay, Expressing

    HG treatment suppresses the expression of PRKAA1 and upregulates levels of miR-137 and IL-6. HTR-8/SVneo trophoblastic cells were treated with HG (25 mmol/l) medium for 24 h, with a control group cultured in normal medium (5 mmol/l). Expression levels of miR-137 and PRKAA1 were detected using reverse transcription-quantitative polymerase chain reaction analysis, with hsa-u6 and β-actin as internal controls. Protein levels of PRKAA1 (~62 kDa) and IL-6 were measured using western blot analysis and ELISA, respectively, and the relative density of PRKAA1 was determined using Image Pro Plus 6.0 software. (A) HG intervention suppressed the expression of PRKAA1 in HTR-8/SVneo cells. (B) miR-137 expression levels were elevated in HG-exposed HTR-8/SVneo cells. (C) Protein level of PRKAA1 was decreased in HG-exposed HTR-8/SVneo cells. (D) Results of western blot analysis. (E) HG treatment promoted the secretion of IL-6 in HTR-8/SVneo cells. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Journal: International Journal of Molecular Medicine

    Article Title: High glucose suppresses the viability and proliferation of HTR-8/SVneo cells through regulation of the miR-137/PRKAA1/IL-6 axis

    doi: 10.3892/ijmm.2018.3686

    Figure Lengend Snippet: HG treatment suppresses the expression of PRKAA1 and upregulates levels of miR-137 and IL-6. HTR-8/SVneo trophoblastic cells were treated with HG (25 mmol/l) medium for 24 h, with a control group cultured in normal medium (5 mmol/l). Expression levels of miR-137 and PRKAA1 were detected using reverse transcription-quantitative polymerase chain reaction analysis, with hsa-u6 and β-actin as internal controls. Protein levels of PRKAA1 (~62 kDa) and IL-6 were measured using western blot analysis and ELISA, respectively, and the relative density of PRKAA1 was determined using Image Pro Plus 6.0 software. (A) HG intervention suppressed the expression of PRKAA1 in HTR-8/SVneo cells. (B) miR-137 expression levels were elevated in HG-exposed HTR-8/SVneo cells. (C) Protein level of PRKAA1 was decreased in HG-exposed HTR-8/SVneo cells. (D) Results of western blot analysis. (E) HG treatment promoted the secretion of IL-6 in HTR-8/SVneo cells. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Article Snippet: Rabbit monoclonal antibodies against β-actin (Cell Signaling Technology, Inc., Danvers, MA, USA, cat. no. 8457) and PRKKA1 (Cell Signaling Technology, Inc., cat. no. 2795S) were diluted in primary antibody dilution buffer (Beyotime Institute of Biotechnology) at a ratio of 1:1,000, and the membranes were incubated in the antibody solution at 4°C overnight.

    Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Software

    PRKAA1 is decreased in placental tissues of women with GDM. Western blot analysis was used to measure the protein level of PRKAA1 in placental tissues of women without GDM (n=11) and with GDM (n=11). β-actin (~43 kDa) was used as the internal control. The relative density of PRKAA1 (~62 kDa) was determined with Image Pro Plus version 6.0 software. (A) Representative protein bands of PRKAA1 and β-actin in placental biopsies of five normal pregnant women and five women with GDM. (B) Relative density of PRKAA1 in placental tissues of the normal (n=11) and GDM (n=11) groups. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Journal: International Journal of Molecular Medicine

    Article Title: High glucose suppresses the viability and proliferation of HTR-8/SVneo cells through regulation of the miR-137/PRKAA1/IL-6 axis

    doi: 10.3892/ijmm.2018.3686

    Figure Lengend Snippet: PRKAA1 is decreased in placental tissues of women with GDM. Western blot analysis was used to measure the protein level of PRKAA1 in placental tissues of women without GDM (n=11) and with GDM (n=11). β-actin (~43 kDa) was used as the internal control. The relative density of PRKAA1 (~62 kDa) was determined with Image Pro Plus version 6.0 software. (A) Representative protein bands of PRKAA1 and β-actin in placental biopsies of five normal pregnant women and five women with GDM. (B) Relative density of PRKAA1 in placental tissues of the normal (n=11) and GDM (n=11) groups. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Article Snippet: Rabbit monoclonal antibodies against β-actin (Cell Signaling Technology, Inc., Danvers, MA, USA, cat. no. 8457) and PRKKA1 (Cell Signaling Technology, Inc., cat. no. 2795S) were diluted in primary antibody dilution buffer (Beyotime Institute of Biotechnology) at a ratio of 1:1,000, and the membranes were incubated in the antibody solution at 4°C overnight.

    Techniques: Western Blot, Software

    miR-137 upregulation decreases levels of PRKAA1 in HTR-8/SVneo cells. HTR-8/SVneo cells were infected with lentiviral vectors and miR-137 and PRKAA1 gene expression levels were measured using reverse transcription-quantitative polymerase chain reaction analysis, with hsa-u6 and β-actin as internal controls, respectively. Protein levels of PRKAA1 (~62 kDa) were detected by western blot analysis, and β-actin (~43 kDa) was used as an internal control. (A) miR-137 was upregulated in LV-miR-137 HTR-8/SVneo cells. (B) Gene expression level of PRKAA1 was downregulated in the LV-miR-137 group. (C) Western blot analysis of PRKAA1. (D) Protein level of PRKAA1 was decreased in miR-137-overexpressing HTR-8/SVneo cells. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Journal: International Journal of Molecular Medicine

    Article Title: High glucose suppresses the viability and proliferation of HTR-8/SVneo cells through regulation of the miR-137/PRKAA1/IL-6 axis

    doi: 10.3892/ijmm.2018.3686

    Figure Lengend Snippet: miR-137 upregulation decreases levels of PRKAA1 in HTR-8/SVneo cells. HTR-8/SVneo cells were infected with lentiviral vectors and miR-137 and PRKAA1 gene expression levels were measured using reverse transcription-quantitative polymerase chain reaction analysis, with hsa-u6 and β-actin as internal controls, respectively. Protein levels of PRKAA1 (~62 kDa) were detected by western blot analysis, and β-actin (~43 kDa) was used as an internal control. (A) miR-137 was upregulated in LV-miR-137 HTR-8/SVneo cells. (B) Gene expression level of PRKAA1 was downregulated in the LV-miR-137 group. (C) Western blot analysis of PRKAA1. (D) Protein level of PRKAA1 was decreased in miR-137-overexpressing HTR-8/SVneo cells. Data are expressed as the mean ± standard error of the mean; statistical significance was determined using Student's t-test, * P

    Article Snippet: Rabbit monoclonal antibodies against β-actin (Cell Signaling Technology, Inc., Danvers, MA, USA, cat. no. 8457) and PRKKA1 (Cell Signaling Technology, Inc., cat. no. 2795S) were diluted in primary antibody dilution buffer (Beyotime Institute of Biotechnology) at a ratio of 1:1,000, and the membranes were incubated in the antibody solution at 4°C overnight.

    Techniques: Infection, Expressing, Real-time Polymerase Chain Reaction, Western Blot