primary antibodies against kir6 2  (Alomone Labs)


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    Structured Review

    Alomone Labs primary antibodies against kir6 2
    Distribution and localization of <t>Kir6.2</t> immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.
    Primary Antibodies Against Kir6 2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against kir6 2/product/Alomone Labs
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    primary antibodies against kir6 2 - by Bioz Stars, 2022-10
    94/100 stars

    Images

    1) Product Images from "KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock"

    Article Title: KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-00699-3

    Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.
    Figure Legend Snippet: Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.

    Techniques Used: Staining

    Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P
    Figure Legend Snippet: Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction

    2) Product Images from "KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock"

    Article Title: KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-00699-3

    Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.
    Figure Legend Snippet: Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.

    Techniques Used: Staining

    Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P
    Figure Legend Snippet: Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction

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    Alomone Labs antibodies against kir6 1
    Nico ameliorated LPS-induced ALI and inflammation. (a) Nico increased LPS-induced <t>Kir6.1</t> and Kir6.2 downregulation in the lung. (b, c) Lung sections stained with H E showed severe injury in the LPS group which was attenuated by Nico pretreatment. The data revealed a high score for the LPS-treated group which was decreased in the Nico-pretreated group. (d) Nico pretreatment significantly reduced LPS-induced protein leakage in BALF. (e, f) Nico alleviated LPS-induced increments of MPO activities in BALF and lung homogenate. (g, h) Nico prevented the production of TNF- α and IL-1 β in lung homogenate. Data were shown as mean ± SEM ( n = 6 − 8). Statistically significant differences: ∗ P
    Antibodies Against Kir6 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies against kir6 1/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibodies against kir6 1 - by Bioz Stars, 2022-10
    91/100 stars
      Buy from Supplier

    94
    Alomone Labs primary antibodies against kir6 2
    Distribution and localization of <t>Kir6.2</t> immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.
    Primary Antibodies Against Kir6 2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against kir6 2/product/Alomone Labs
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    primary antibodies against kir6 2 - by Bioz Stars, 2022-10
    94/100 stars
      Buy from Supplier

    Image Search Results


    Nico ameliorated LPS-induced ALI and inflammation. (a) Nico increased LPS-induced Kir6.1 and Kir6.2 downregulation in the lung. (b, c) Lung sections stained with H E showed severe injury in the LPS group which was attenuated by Nico pretreatment. The data revealed a high score for the LPS-treated group which was decreased in the Nico-pretreated group. (d) Nico pretreatment significantly reduced LPS-induced protein leakage in BALF. (e, f) Nico alleviated LPS-induced increments of MPO activities in BALF and lung homogenate. (g, h) Nico prevented the production of TNF- α and IL-1 β in lung homogenate. Data were shown as mean ± SEM ( n = 6 − 8). Statistically significant differences: ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Nicorandil Attenuates LPS-Induced Acute Lung Injury by Pulmonary Endothelial Cell Protection via NF-κB and MAPK Pathways

    doi: 10.1155/2019/4957646

    Figure Lengend Snippet: Nico ameliorated LPS-induced ALI and inflammation. (a) Nico increased LPS-induced Kir6.1 and Kir6.2 downregulation in the lung. (b, c) Lung sections stained with H E showed severe injury in the LPS group which was attenuated by Nico pretreatment. The data revealed a high score for the LPS-treated group which was decreased in the Nico-pretreated group. (d) Nico pretreatment significantly reduced LPS-induced protein leakage in BALF. (e, f) Nico alleviated LPS-induced increments of MPO activities in BALF and lung homogenate. (g, h) Nico prevented the production of TNF- α and IL-1 β in lung homogenate. Data were shown as mean ± SEM ( n = 6 − 8). Statistically significant differences: ∗ P

    Article Snippet: Then, the transferred membranes were incubated with primary antibodies against Kir6.1 (Alomone Labs, Jerusalem, Israel), Kir6.2 (Abcam), NF-κ B p-p65/p65, p-iκ B-α /iκ B-α , p-p38/p38, p-ERK/ERK, p-JNK/JNK, intercellular adhesion molecule-1 (ICAM-1), cleaved-caspase-3 (c-caspase-3), caspase-9 (1 : 1000, Cell Signaling Technology), endothelial nitric oxide synthase (eNOS) (1 : 1000, Santa Cruz), inducible nitric oxide synthase (iNOS) (1 : 1000, Millipore), CCAAT/enhancer-binding protein homologous protein (CHOP), vascular cell adhesion molecule-1 (VCAM-1), VE-cadherin, Nox4 (1 : 1000), MnSOD (1 : 5000, Abcam), and β -actin (1 : 5000, Proteintech, Rosemont, USA) overnight.

    Techniques: Staining

    Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.

    Journal: Scientific Reports

    Article Title: KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock

    doi: 10.1038/s41598-017-00699-3

    Figure Lengend Snippet: Distribution and localization of Kir6.2 immunoreactivity (ir) in the SCN. ( A ) Distribution of the Kir6.2 (A1–3) and AVP (A4-6) immunoreactivity in the rostral (A1, A4), middle (A2, A5), and caudal (A3, A6) sections of the SCN. Scale bar: 200 µm. ( B ) Selective colocalization of Kir6.2-ir with AVP-ir (B1–3) but not with VIP-ir (B4–6). Note the colocalization of Kir6.2-ir and AVP-ir in and around the soma (B2) and in varicosities along the process (marked by arrowheads, B3). Note the reciprocal apposition of VIP-ir bouton-like swellings against a Kir6.2-ir soma (B5) and Kir6.2-ir bouton-like swellings against VIP-ir somata (B6). Note also bouton-like swellings (yellow) double-stained with Kir6.2 (green) and AVP (red) apposing Hoechst-stained cells (blue) in the ventrolateral region of the SCN (B7). Scale bar: 100 µm (B1, B4); 10 µm (B2, B3, B5–7). ( C ) Lack of colocalization of Kir6.2-ir with markers for three afferent inputs SERT-ir (C1, C2), vGluT2-ir (C3, C4), NPY-ir (C5, C6). Insets: co-distribution of SERT-ir, vGluT2-ir, and NPY-ir with Kir6.2-stained somata in the ventromedial region of the mid-SCN section. Scale bar: 100 µm (C1, C3, C5); 10 µm (C2, C4, C6, insets). OC: optic chiasm. 3 V: third ventricle. Asterisks mark Hoechst-stained nuclei.

    Article Snippet: For immunohistochemical staining, sections (20 µm) were treated with 0.3% H2 O2 for 15 min to quench endogenous peroxidase, and then incubated overnight at 4 °C in PBS containing 2% serum, 0.3% Triton X-100, and primary antibodies against Kir6.2 (rabbit anti-Kir6.2; 1:5000; APC-020; Alomone Labs, Jerusalem, Israel) and AVP (rabbit anti-AVP; 1:3000; AB1565; Millipore, Temecula, CA, USA) .

    Techniques: Staining

    Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P

    Journal: Scientific Reports

    Article Title: KATP Channels Mediate Differential Metabolic Responses to Glucose Shortage of the Dorsomedial and Ventrolateral Oscillators in the Central Clock

    doi: 10.1038/s41598-017-00699-3

    Figure Lengend Snippet: Kir6.2/SUR1 combination of K ATP channels in the SCN neurones. ( A ) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. ( B ) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz ( n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz ( n = 17). * P

    Article Snippet: For immunohistochemical staining, sections (20 µm) were treated with 0.3% H2 O2 for 15 min to quench endogenous peroxidase, and then incubated overnight at 4 °C in PBS containing 2% serum, 0.3% Triton X-100, and primary antibodies against Kir6.2 (rabbit anti-Kir6.2; 1:5000; APC-020; Alomone Labs, Jerusalem, Israel) and AVP (rabbit anti-AVP; 1:3000; AB1565; Millipore, Temecula, CA, USA) .

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction

    Chromatin immunoprecipitation (ChIP) assay demonstrating binding of HIF-2α to the promoter region of Kir6.2 gene in opioid-treated MAH cells. Note hypoxia response element (HRE) within the promoter region of rat and mouse Kir6.2 gene ( A , top ); the HIF core site (GCGTG) spans nucleotides −1087 to −1083 and HIF ancillary site (CACAG) spans nucleotides −1065 to −1061. Lysates from untreated control (Untr) and opioid-treated wt, shHIF2α, and ScCont MAH cells were subjected to ChIP assay using a HIF-2α polyclonal antibody ( A , bottom ). PCR analysis was performed using a primer pair designed to span the putative HRE upstream of the promoter region. Technical controls include a ChIP performed using nonspecific IgG monoclonal antibody (IgG) and a starting material control (Input). B : vascular endothelial growth factor (VEGF) mRNA expression as determined by QPCR analysis in control (untreated) and opioid-treated MAH cells ( n = 3). Data are expressed as means ± SE for three independent experiments for each group, * P

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Chronic opioids regulate KATP channel subunit Kir6.2 and carbonic anhydrase I and II expression in rat adrenal chromaffin cells via HIF-2α and protein kinase A

    doi: 10.1152/ajpcell.00135.2014

    Figure Lengend Snippet: Chromatin immunoprecipitation (ChIP) assay demonstrating binding of HIF-2α to the promoter region of Kir6.2 gene in opioid-treated MAH cells. Note hypoxia response element (HRE) within the promoter region of rat and mouse Kir6.2 gene ( A , top ); the HIF core site (GCGTG) spans nucleotides −1087 to −1083 and HIF ancillary site (CACAG) spans nucleotides −1065 to −1061. Lysates from untreated control (Untr) and opioid-treated wt, shHIF2α, and ScCont MAH cells were subjected to ChIP assay using a HIF-2α polyclonal antibody ( A , bottom ). PCR analysis was performed using a primer pair designed to span the putative HRE upstream of the promoter region. Technical controls include a ChIP performed using nonspecific IgG monoclonal antibody (IgG) and a starting material control (Input). B : vascular endothelial growth factor (VEGF) mRNA expression as determined by QPCR analysis in control (untreated) and opioid-treated MAH cells ( n = 3). Data are expressed as means ± SE for three independent experiments for each group, * P

    Article Snippet: Membranes were then washed and incubated with either primary rabbit polyclonal antibody against Kir6.2 (catalog no. APC-020; 1:1,000 dilution; Alomone Labs, Jerusalem, Israel), rabbit polyclonal anti-human CAI antibody (catalog no. ; 1:1,000 dilution; Abcam, Cambridge, MA), sheep polyclonal anti-human CAII antibody (catalog no. AHP 206; 1:1,000 dilution; AbD Serotec, Kidlington, UK), HIF-1α mouse monoclonal antibody (catalog no. NB 100–105; 1:1,000 dilution; Novus Biologicals, Littleton, CO), or HIF-2α rabbit polyclonal antibody (catalog no. NB 100–122; 1:1,000 dilution; Novus Biologicals), primary rabbit monoclonal β-actin antibody as loading control for cytoplasmic extracts (1:10,000 dilution; Millipore, Billerica, MA), or primary rabbit polyclonal TATA-binding protein antibody as a loading control for nuclear extracts (1:25,000 dilution; Santa Cruz) at 4°C overnight.

    Techniques: Chromatin Immunoprecipitation, Binding Assay, Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction