kt 5823  (Alomone Labs)


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    Name:
    KT5823
    Description:
    KT5823 is a selective and potent inhibitor of PKG IC50 234 nM
    Catalog Number:
    K-250
    Price:
    300.0
    Category:
    Small Molecule
    Source:
    Nocardiopsis sp. (soil fungi).
    Applications:
    0
    Purity:
    >98%
    Size:
    1 Vials containing 50 mcg each
    Format:
    Lyophilized/solid.
    Formula:
    C29H25N3O5
    Molecular Weight:
    495
    Molecule Name:
    (9S,10R,12R)-2,3,9,10,11,12-Hexahydro-10-methoxy-2,9-di methyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-k l]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid, methyl ester.
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    Structured Review

    Alomone Labs kt 5823
    KT5823
    KT5823 is a selective and potent inhibitor of PKG IC50 234 nM
    https://www.bioz.com/result/kt 5823/product/Alomone Labs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kt 5823 - by Bioz Stars, 2021-09
    85/100 stars

    Images

    1) Product Images from "Low Doses of Bisphenol A and Diethylstilbestrol Impair Ca2+ Signals in Pancreatic ?-Cells through a Nonclassical Membrane Estrogen Receptor within Intact Islets of Langerhans"

    Article Title: Low Doses of Bisphenol A and Diethylstilbestrol Impair Ca2+ Signals in Pancreatic ?-Cells through a Nonclassical Membrane Estrogen Receptor within Intact Islets of Langerhans

    Journal: Environmental Health Perspectives

    doi: 10.1289/ehp.8002

    Effects of BPA on [Ca 2+ ] i oscillations through a PKG-mediated mechanism. ( A ) Low-glucose–induced [Ca 2+ ] i oscillations blocked by 1 nM BPA. ( B ) Frequency of [Ca 2+ ] i oscillations were not reduced by BPA in an islet from the same preparation and maintained in the same conditions but pretreated with and exposed to the PKG inhibitor KT-5823 (1 μM). ( C ) Mean frequency values of 0.5 mM glucose before application of either BPA (G1) or E 2 (G2), in the presence of 1 nM BPA or 1 nM E 2 , as in ( A ) , or in the presence of 1 μM KT-5823 plus 0.5 mM glucose (KT); KT plus 1 nM BPA (BPA + KT), and KT plus 1 nM 17β-E 2 (E 2 + KT) as in ( B ). Results are representative of at least 12 cells from nine different islets, expressed as mean ± SE. * p
    Figure Legend Snippet: Effects of BPA on [Ca 2+ ] i oscillations through a PKG-mediated mechanism. ( A ) Low-glucose–induced [Ca 2+ ] i oscillations blocked by 1 nM BPA. ( B ) Frequency of [Ca 2+ ] i oscillations were not reduced by BPA in an islet from the same preparation and maintained in the same conditions but pretreated with and exposed to the PKG inhibitor KT-5823 (1 μM). ( C ) Mean frequency values of 0.5 mM glucose before application of either BPA (G1) or E 2 (G2), in the presence of 1 nM BPA or 1 nM E 2 , as in ( A ) , or in the presence of 1 μM KT-5823 plus 0.5 mM glucose (KT); KT plus 1 nM BPA (BPA + KT), and KT plus 1 nM 17β-E 2 (E 2 + KT) as in ( B ). Results are representative of at least 12 cells from nine different islets, expressed as mean ± SE. * p

    Techniques Used:

    Effects of 8Br-cGMP on EDCs and E 2 via PKG. ( A ) Exposure to 10 μM 8Br-cGMP dramatically reduces the frequency of low-glucose–induced [Ca 2+ ] i oscillations. ( B ) After incubation with the specific PKG inhibitor KT-5823 (1 μM), 8Br-cGMP fails to evoke the marked reduction in [Ca 2+ ] i oscillations shown in ( A ). ( C ) Mean frequency values collected in the presence of 0.5 mM glucose (G), 8Br-cGMP plus 0.5 mM glucose (8Br-cGMP), KT-5823 (KT), and 8Br-cGMP plus KT-5823 (8Br-cGMP + KT). Results are representative of at least five cells in four different islets, expressed as mean ± SE.
    Figure Legend Snippet: Effects of 8Br-cGMP on EDCs and E 2 via PKG. ( A ) Exposure to 10 μM 8Br-cGMP dramatically reduces the frequency of low-glucose–induced [Ca 2+ ] i oscillations. ( B ) After incubation with the specific PKG inhibitor KT-5823 (1 μM), 8Br-cGMP fails to evoke the marked reduction in [Ca 2+ ] i oscillations shown in ( A ). ( C ) Mean frequency values collected in the presence of 0.5 mM glucose (G), 8Br-cGMP plus 0.5 mM glucose (8Br-cGMP), KT-5823 (KT), and 8Br-cGMP plus KT-5823 (8Br-cGMP + KT). Results are representative of at least five cells in four different islets, expressed as mean ± SE.

    Techniques Used: Incubation

    Related Articles

    other:

    Article Title: Expression of olfactory-type cyclic nucleotide-gated channels in rat cortical astrocytes.
    Article Snippet: Cyclic nucleotide-gated (CNG) channels are nonselective cation channels activated by cyclic AMP (cAMP) or cyclic GMP (cGMP).. They were originally identified in retinal and olfactory receptors, but evidence has also emerged for their expression in several mammalian brain areas.

    Article Title: Low Doses of Bisphenol A and Diethylstilbestrol Impair Ca2+ Signals in Pancreatic ?-Cells through a Nonclassical Membrane Estrogen Receptor within Intact Islets of Langerhans
    Article Snippet: Materials.We obtained Fluo-3 AM from Molecular Probes Inc. (Leiden, the Netherlands); ICI182,780 and 1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one (ODQ) from Tocris Cookson Ltd. (Avonmouth, UK); and KT-5823 from Alomone Labs (Jerusalem, Israel).

    Article Title: Staurosporine induces lamellipodial widening in locomoting fish keratocytes by abolishing the gradient from radial extension of leading edge
    Article Snippet: Protein-serine/threonine kinase inhibitorsStaurosporine, KT5720, and KT5823 were products of Aromone Labs (Jerusalem, Israel), and Ro 31-8220, SB-415286, KN-93, K-252a, and K-252b were purchased from Biomol Research Laboratories Inc. (Plymouth Meeting, PA, USA), while ML-7 and Y-27632 were from EMD Biosciences, Inc (Merck KGaA, Darmstadt, Germany).

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    Alomone Labs kt 5823
    Effects of BPA on [Ca 2+ ] i oscillations through a PKG-mediated mechanism. ( A ) Low-glucose–induced [Ca 2+ ] i oscillations blocked by 1 nM BPA. ( B ) Frequency of [Ca 2+ ] i oscillations were not reduced by BPA in an islet from the same preparation and maintained in the same conditions but pretreated with and exposed to the PKG inhibitor <t>KT-5823</t> (1 μM). ( C ) Mean frequency values of 0.5 mM glucose before application of either BPA (G1) or E 2 (G2), in the presence of 1 nM BPA or 1 nM E 2 , as in ( A ) , or in the presence of 1 μM KT-5823 plus 0.5 mM glucose (KT); KT plus 1 nM BPA (BPA + KT), and KT plus 1 nM 17β-E 2 (E 2 + KT) as in ( B ). Results are representative of at least 12 cells from nine different islets, expressed as mean ± SE. * p
    Kt 5823, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kt 5823/product/Alomone Labs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kt 5823 - by Bioz Stars, 2021-09
    85/100 stars
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    86
    alomone labs mouse recombinant il 6
    Bile duct staining with CK19 (A–C) and cell proliferation analysis by Ki-67 staining (D, E) were performed 2 weeks after BDL mice. A group of mice received recombinant mouse <t>IL-6</t> (IL-6) or the vehicle only (vehicle) for both sham and BDL surgeries. (A) Representative CK19 staining image for each group. Scale bar: 50 μm. (D, E) Count of CK19 + duct (D; with visible lumen) and nonduct (E; no lumen) per portal area in a blinded manner. The following were the number of animals per group: C57BL/6 sham (7), C57BL/6 sham + IL-6 (3), C57BL/6 BDL (9), C57BL/6 BDL + IL-6 (3), A2bAR −/− sham (7), A2bAR −/− sham + IL-6 (3), A2bAR −/− BDL (13), and A2bAR −/− BDL + IL-6 (3). Between 3 and 5 100× magnification fields were count per animal. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to A2bAR −/− BDL vehicle. (D) Representative image of Ki-67 staining for BDL-operated animal with or without recmIL-6 injection. Scale bar: 50 μm. (E) Analysis of the percent of Ki-67 + pixels on five different 100× magnification fields for three animals per group. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to C57BL/6 BDL IL-6.
    Mouse Recombinant Il 6, supplied by alomone labs, 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/mouse recombinant il 6/product/alomone labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse recombinant il 6 - by Bioz Stars, 2021-09
    86/100 stars
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    Effects of BPA on [Ca 2+ ] i oscillations through a PKG-mediated mechanism. ( A ) Low-glucose–induced [Ca 2+ ] i oscillations blocked by 1 nM BPA. ( B ) Frequency of [Ca 2+ ] i oscillations were not reduced by BPA in an islet from the same preparation and maintained in the same conditions but pretreated with and exposed to the PKG inhibitor KT-5823 (1 μM). ( C ) Mean frequency values of 0.5 mM glucose before application of either BPA (G1) or E 2 (G2), in the presence of 1 nM BPA or 1 nM E 2 , as in ( A ) , or in the presence of 1 μM KT-5823 plus 0.5 mM glucose (KT); KT plus 1 nM BPA (BPA + KT), and KT plus 1 nM 17β-E 2 (E 2 + KT) as in ( B ). Results are representative of at least 12 cells from nine different islets, expressed as mean ± SE. * p

    Journal: Environmental Health Perspectives

    Article Title: Low Doses of Bisphenol A and Diethylstilbestrol Impair Ca2+ Signals in Pancreatic ?-Cells through a Nonclassical Membrane Estrogen Receptor within Intact Islets of Langerhans

    doi: 10.1289/ehp.8002

    Figure Lengend Snippet: Effects of BPA on [Ca 2+ ] i oscillations through a PKG-mediated mechanism. ( A ) Low-glucose–induced [Ca 2+ ] i oscillations blocked by 1 nM BPA. ( B ) Frequency of [Ca 2+ ] i oscillations were not reduced by BPA in an islet from the same preparation and maintained in the same conditions but pretreated with and exposed to the PKG inhibitor KT-5823 (1 μM). ( C ) Mean frequency values of 0.5 mM glucose before application of either BPA (G1) or E 2 (G2), in the presence of 1 nM BPA or 1 nM E 2 , as in ( A ) , or in the presence of 1 μM KT-5823 plus 0.5 mM glucose (KT); KT plus 1 nM BPA (BPA + KT), and KT plus 1 nM 17β-E 2 (E 2 + KT) as in ( B ). Results are representative of at least 12 cells from nine different islets, expressed as mean ± SE. * p

    Article Snippet: Materials.We obtained Fluo-3 AM from Molecular Probes Inc. (Leiden, the Netherlands); ICI182,780 and 1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one (ODQ) from Tocris Cookson Ltd. (Avonmouth, UK); and KT-5823 from Alomone Labs (Jerusalem, Israel).

    Techniques:

    Effects of 8Br-cGMP on EDCs and E 2 via PKG. ( A ) Exposure to 10 μM 8Br-cGMP dramatically reduces the frequency of low-glucose–induced [Ca 2+ ] i oscillations. ( B ) After incubation with the specific PKG inhibitor KT-5823 (1 μM), 8Br-cGMP fails to evoke the marked reduction in [Ca 2+ ] i oscillations shown in ( A ). ( C ) Mean frequency values collected in the presence of 0.5 mM glucose (G), 8Br-cGMP plus 0.5 mM glucose (8Br-cGMP), KT-5823 (KT), and 8Br-cGMP plus KT-5823 (8Br-cGMP + KT). Results are representative of at least five cells in four different islets, expressed as mean ± SE.

    Journal: Environmental Health Perspectives

    Article Title: Low Doses of Bisphenol A and Diethylstilbestrol Impair Ca2+ Signals in Pancreatic ?-Cells through a Nonclassical Membrane Estrogen Receptor within Intact Islets of Langerhans

    doi: 10.1289/ehp.8002

    Figure Lengend Snippet: Effects of 8Br-cGMP on EDCs and E 2 via PKG. ( A ) Exposure to 10 μM 8Br-cGMP dramatically reduces the frequency of low-glucose–induced [Ca 2+ ] i oscillations. ( B ) After incubation with the specific PKG inhibitor KT-5823 (1 μM), 8Br-cGMP fails to evoke the marked reduction in [Ca 2+ ] i oscillations shown in ( A ). ( C ) Mean frequency values collected in the presence of 0.5 mM glucose (G), 8Br-cGMP plus 0.5 mM glucose (8Br-cGMP), KT-5823 (KT), and 8Br-cGMP plus KT-5823 (8Br-cGMP + KT). Results are representative of at least five cells in four different islets, expressed as mean ± SE.

    Article Snippet: Materials.We obtained Fluo-3 AM from Molecular Probes Inc. (Leiden, the Netherlands); ICI182,780 and 1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one (ODQ) from Tocris Cookson Ltd. (Avonmouth, UK); and KT-5823 from Alomone Labs (Jerusalem, Israel).

    Techniques: Incubation

    Bile duct staining with CK19 (A–C) and cell proliferation analysis by Ki-67 staining (D, E) were performed 2 weeks after BDL mice. A group of mice received recombinant mouse IL-6 (IL-6) or the vehicle only (vehicle) for both sham and BDL surgeries. (A) Representative CK19 staining image for each group. Scale bar: 50 μm. (D, E) Count of CK19 + duct (D; with visible lumen) and nonduct (E; no lumen) per portal area in a blinded manner. The following were the number of animals per group: C57BL/6 sham (7), C57BL/6 sham + IL-6 (3), C57BL/6 BDL (9), C57BL/6 BDL + IL-6 (3), A2bAR −/− sham (7), A2bAR −/− sham + IL-6 (3), A2bAR −/− BDL (13), and A2bAR −/− BDL + IL-6 (3). Between 3 and 5 100× magnification fields were count per animal. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to A2bAR −/− BDL vehicle. (D) Representative image of Ki-67 staining for BDL-operated animal with or without recmIL-6 injection. Scale bar: 50 μm. (E) Analysis of the percent of Ki-67 + pixels on five different 100× magnification fields for three animals per group. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to C57BL/6 BDL IL-6.

    Journal: Gene Expression

    Article Title: The Cholangiocyte Adenosine–IL-6 Axis Regulates Survival During Biliary Cirrhosis

    doi: 10.3727/105221617X15042723767876

    Figure Lengend Snippet: Bile duct staining with CK19 (A–C) and cell proliferation analysis by Ki-67 staining (D, E) were performed 2 weeks after BDL mice. A group of mice received recombinant mouse IL-6 (IL-6) or the vehicle only (vehicle) for both sham and BDL surgeries. (A) Representative CK19 staining image for each group. Scale bar: 50 μm. (D, E) Count of CK19 + duct (D; with visible lumen) and nonduct (E; no lumen) per portal area in a blinded manner. The following were the number of animals per group: C57BL/6 sham (7), C57BL/6 sham + IL-6 (3), C57BL/6 BDL (9), C57BL/6 BDL + IL-6 (3), A2bAR −/− sham (7), A2bAR −/− sham + IL-6 (3), A2bAR −/− BDL (13), and A2bAR −/− BDL + IL-6 (3). Between 3 and 5 100× magnification fields were count per animal. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to A2bAR −/− BDL vehicle. (D) Representative image of Ki-67 staining for BDL-operated animal with or without recmIL-6 injection. Scale bar: 50 μm. (E) Analysis of the percent of Ki-67 + pixels on five different 100× magnification fields for three animals per group. #Significant when compared to C57BL/6 BDL vehicle; *significant when compared to C57BL/6 BDL IL-6.

    Article Snippet: For some experiments, A2bAR−/− and C57BL/6 mice were injected subcutaneously with 250 ng/g of body weight of mouse recombinant IL-6 (recmIL-6; BioLegend, San Diego, CA, USA) or vehicle 15 min prior to the surgery and once a week for the duration of the experiments.

    Techniques: Staining, Mouse Assay, Recombinant, Injection

    Second messenger alterations and control of IL-6 release mediated by adenosine/A2bAR. Intracellular cAMP concentrations (A, B) were analyzed by ELISA. H69 cells were stimulated with 100 μM adenosine for 2 h in 96-well plates (5,000 cells per well) in H69 complete media in the presence of A2bAR antagonist (A) MRS-1754 (1 μM; n = 3, performed in triplicates) or (B) 72 h after transfection with an A2bAR-specific siRNA ( n = 4, performed in triplicate). #Significant when compared to the untreated cells; *significant when compared to vehicle (A) or nontarget siRNA (B). (C, D) Intracellular Ca 2+ mobilization was observed by confocal microscopy. Glass coverslip-plated, fluo-4/AM-loaded H69 cells were perifused with HEPES buffer alone or containing 500 μM adenosine. ATP (100 μM) was used as a positive control. Changes in fluorescence intensity were recorded with a Zeiss 510 confocal microscope. (C) Relative fluorescence intensity of individual cells; 29 individual cells were analyzed. (D) Representative image of the dye intensity variation during the course of the experiment. Scale bar: 50 μm. (E) H69 cells were stimulated by 100 μM of adenosine for 2 h in six-well plates (150,000 cells per well) in H69 complete media. RNA was extracted, and IL-6 mRNA expression was determined by qPCR. Cells were preincubated with BAPTA/AM (50 μM) or cAMPs-RP (100 μM) 30 min prior to the stimulation ( n = 3 for cAMPs-RP and n = 7 for BAPTA/AM). (F) H69 cells were stimulated by 100 μM adenosine for 4 h in 48-well plates (30,000 cells per well) in H69 complete media. Cells were preincubated with BAPTA/AM (50 μM) or cAMPs-RP (100 μM) 30 min prior to the stimulation. Supernatants were collected and centrifuged, and IL-6 content was analyzed by ELISA ( n = 3, performed in triplicate). #Significant when compared to unstimulated cells; *significant when compared to vehicle. (G) H69 cells were stimulated by 100 μM of 8-pCPT-2-O-Me-cAMP-AM (8-pCPT) or adenosine for 4 h in 48-well plates (30,000 cells per well) in H69 complete media. Supernatants were collected and centrifuged, and IL-6 content was analyzed by ELISA ( n = 3, performed in triplicate).

    Journal: Gene Expression

    Article Title: The Cholangiocyte Adenosine–IL-6 Axis Regulates Survival During Biliary Cirrhosis

    doi: 10.3727/105221617X15042723767876

    Figure Lengend Snippet: Second messenger alterations and control of IL-6 release mediated by adenosine/A2bAR. Intracellular cAMP concentrations (A, B) were analyzed by ELISA. H69 cells were stimulated with 100 μM adenosine for 2 h in 96-well plates (5,000 cells per well) in H69 complete media in the presence of A2bAR antagonist (A) MRS-1754 (1 μM; n = 3, performed in triplicates) or (B) 72 h after transfection with an A2bAR-specific siRNA ( n = 4, performed in triplicate). #Significant when compared to the untreated cells; *significant when compared to vehicle (A) or nontarget siRNA (B). (C, D) Intracellular Ca 2+ mobilization was observed by confocal microscopy. Glass coverslip-plated, fluo-4/AM-loaded H69 cells were perifused with HEPES buffer alone or containing 500 μM adenosine. ATP (100 μM) was used as a positive control. Changes in fluorescence intensity were recorded with a Zeiss 510 confocal microscope. (C) Relative fluorescence intensity of individual cells; 29 individual cells were analyzed. (D) Representative image of the dye intensity variation during the course of the experiment. Scale bar: 50 μm. (E) H69 cells were stimulated by 100 μM of adenosine for 2 h in six-well plates (150,000 cells per well) in H69 complete media. RNA was extracted, and IL-6 mRNA expression was determined by qPCR. Cells were preincubated with BAPTA/AM (50 μM) or cAMPs-RP (100 μM) 30 min prior to the stimulation ( n = 3 for cAMPs-RP and n = 7 for BAPTA/AM). (F) H69 cells were stimulated by 100 μM adenosine for 4 h in 48-well plates (30,000 cells per well) in H69 complete media. Cells were preincubated with BAPTA/AM (50 μM) or cAMPs-RP (100 μM) 30 min prior to the stimulation. Supernatants were collected and centrifuged, and IL-6 content was analyzed by ELISA ( n = 3, performed in triplicate). #Significant when compared to unstimulated cells; *significant when compared to vehicle. (G) H69 cells were stimulated by 100 μM of 8-pCPT-2-O-Me-cAMP-AM (8-pCPT) or adenosine for 4 h in 48-well plates (30,000 cells per well) in H69 complete media. Supernatants were collected and centrifuged, and IL-6 content was analyzed by ELISA ( n = 3, performed in triplicate).

    Article Snippet: For some experiments, A2bAR−/− and C57BL/6 mice were injected subcutaneously with 250 ng/g of body weight of mouse recombinant IL-6 (recmIL-6; BioLegend, San Diego, CA, USA) or vehicle 15 min prior to the surgery and once a week for the duration of the experiments.

    Techniques: Enzyme-linked Immunosorbent Assay, Transfection, Confocal Microscopy, Positive Control, Fluorescence, Microscopy, Expressing, Real-time Polymerase Chain Reaction

    Effect of A2bAR knockout on survival and fibrosis after bile duct ligation (BDL). BDL was performed on 10- to 12-week-old male C57BL/6 or A2bAR −/− mice. (A–D) A group of mice received weekly subcutaneous injection of recombinant mouse IL-6 (IL-6; n = 13 for both background), and the other group received vehicle only ( n = 8 for both background). (A) Kaplan–Meier survival graph. The black line represents control C57BL/6 mice, and the blue line corresponds to A2bAR −/− mice. Full lines are vehicle only, and dotted lines represent mice that received recmIL-6 injections. *Significant when compared to C57BL/6 mice; #significant when compared to vehicle injection. (B) Mason trichrome staining representative of each group at the median time of survival. Scale bar: 100 μm. (C) METAVIR fibrosis score analysis of the Mason trichrome staining performed in a blinded manner. (D) Bile infarcts were counted on a blinded manner on low-power field hematoxylin and eosin staining of each individual mouse. (E–H) BDL or sham operation was performed on 10- to 12-week-old male C57BL/6 ( n = 9 and 7, respectively) or A2bAR −/− mice ( n = 13 and 7, respectively). Mice were sacrificed 2 weeks after surgery. (E) Representative picture of hematoxylin and eosin (H E), Pico Sirius red (PSR), and Mason trichrome staining for each group of mice. Scale bar: 100 μm. (F) METAVIR fibrosis score analysis of the Mason trichrome staining performed in a blinded manner. (G) Total liver RNA was extracted, and relative IL-6 mRNA was assessed by qPCR. GAPDH -specific probe was used as control. C57BL/6 and A2bAR −/− sham operated levels were, respectively, used as reference for the BDL group. (H) Serum was obtained from sham and BDL-operated mice, and total serum IL-6 was assessed by ELISA.

    Journal: Gene Expression

    Article Title: The Cholangiocyte Adenosine–IL-6 Axis Regulates Survival During Biliary Cirrhosis

    doi: 10.3727/105221617X15042723767876

    Figure Lengend Snippet: Effect of A2bAR knockout on survival and fibrosis after bile duct ligation (BDL). BDL was performed on 10- to 12-week-old male C57BL/6 or A2bAR −/− mice. (A–D) A group of mice received weekly subcutaneous injection of recombinant mouse IL-6 (IL-6; n = 13 for both background), and the other group received vehicle only ( n = 8 for both background). (A) Kaplan–Meier survival graph. The black line represents control C57BL/6 mice, and the blue line corresponds to A2bAR −/− mice. Full lines are vehicle only, and dotted lines represent mice that received recmIL-6 injections. *Significant when compared to C57BL/6 mice; #significant when compared to vehicle injection. (B) Mason trichrome staining representative of each group at the median time of survival. Scale bar: 100 μm. (C) METAVIR fibrosis score analysis of the Mason trichrome staining performed in a blinded manner. (D) Bile infarcts were counted on a blinded manner on low-power field hematoxylin and eosin staining of each individual mouse. (E–H) BDL or sham operation was performed on 10- to 12-week-old male C57BL/6 ( n = 9 and 7, respectively) or A2bAR −/− mice ( n = 13 and 7, respectively). Mice were sacrificed 2 weeks after surgery. (E) Representative picture of hematoxylin and eosin (H E), Pico Sirius red (PSR), and Mason trichrome staining for each group of mice. Scale bar: 100 μm. (F) METAVIR fibrosis score analysis of the Mason trichrome staining performed in a blinded manner. (G) Total liver RNA was extracted, and relative IL-6 mRNA was assessed by qPCR. GAPDH -specific probe was used as control. C57BL/6 and A2bAR −/− sham operated levels were, respectively, used as reference for the BDL group. (H) Serum was obtained from sham and BDL-operated mice, and total serum IL-6 was assessed by ELISA.

    Article Snippet: For some experiments, A2bAR−/− and C57BL/6 mice were injected subcutaneously with 250 ng/g of body weight of mouse recombinant IL-6 (recmIL-6; BioLegend, San Diego, CA, USA) or vehicle 15 min prior to the surgery and once a week for the duration of the experiments.

    Techniques: Knock-Out, Ligation, Mouse Assay, Injection, Recombinant, Staining, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay