Structured Review

Cayman Chemical nifedipine
The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM <t>nifedipine</t> (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p
Nifedipine, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/nifedipine/product/Cayman Chemical
Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
nifedipine - by Bioz Stars, 2020-09
92/100 stars

Images

1) Product Images from "Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension"

Article Title: Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension

Journal: PLoS ONE

doi: 10.1371/journal.pone.0195780

The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p
Figure Legend Snippet: The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

Techniques Used: Standard Deviation, Incubation

2) Product Images from "Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension"

Article Title: Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension

Journal: PLoS ONE

doi: 10.1371/journal.pone.0195780

The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p
Figure Legend Snippet: The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

Techniques Used: Standard Deviation, Incubation

3) Product Images from "Drug repositioning in epilepsy reveals novel antiseizure candidates"

Article Title: Drug repositioning in epilepsy reveals novel antiseizure candidates

Journal: Annals of Clinical and Translational Neurology

doi: 10.1002/acn3.703

Larval zebrafish behavioral assays. (A) Doxycycline behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 7–8). In the first half‐hour behavioral assay (Test 1), larvae were treated with vehicle (−) or doxycycline (Dox). In the second half‐hour behavioral assay (Test 2), larvae were treated with vehicle, Dox, or PTZ . Movement greater than 20 mm/sec is shown in A’. (B) Metformin behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 30–66). In the first half‐hour behavioral assay, larvae were treated with vehicle or metformin (Met). In the second half‐hour behavioral assay, larvae were treated with vehicle, Met, or PTZ . Movement greater than 20 mm/sec is shown in B’. (C) Nifedipine behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38). In the first half‐hour behavioral assay, larvae were treated with vehicle or nifedipine (Nif). In the second half‐hour behavioral assay, larvae were treated with vehicle, Nif, or PTZ . Movement greater than 20 mm/sec is shown in C’. (D) Pyrantel tartrate behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38–40). In the first half‐hour behavioral assay, larvae were treated with vehicle or pyrantel tartrate (Pyr). In the second half‐hour behavioral assay, larvae were treated with vehicle, Pyr, or PTZ . Movement greater than 20 mm/sec is shown in D’. Bars represent mean ± SEM . **** P ‐value
Figure Legend Snippet: Larval zebrafish behavioral assays. (A) Doxycycline behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 7–8). In the first half‐hour behavioral assay (Test 1), larvae were treated with vehicle (−) or doxycycline (Dox). In the second half‐hour behavioral assay (Test 2), larvae were treated with vehicle, Dox, or PTZ . Movement greater than 20 mm/sec is shown in A’. (B) Metformin behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 30–66). In the first half‐hour behavioral assay, larvae were treated with vehicle or metformin (Met). In the second half‐hour behavioral assay, larvae were treated with vehicle, Met, or PTZ . Movement greater than 20 mm/sec is shown in B’. (C) Nifedipine behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38). In the first half‐hour behavioral assay, larvae were treated with vehicle or nifedipine (Nif). In the second half‐hour behavioral assay, larvae were treated with vehicle, Nif, or PTZ . Movement greater than 20 mm/sec is shown in C’. (D) Pyrantel tartrate behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38–40). In the first half‐hour behavioral assay, larvae were treated with vehicle or pyrantel tartrate (Pyr). In the second half‐hour behavioral assay, larvae were treated with vehicle, Pyr, or PTZ . Movement greater than 20 mm/sec is shown in D’. Bars represent mean ± SEM . **** P ‐value

Techniques Used: Behavioral Assay, Size-exclusion Chromatography

4) Product Images from "Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit"

Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit

Journal: PLoS ONE

doi: 10.1371/journal.pone.0202039

CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P
Figure Legend Snippet: CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P

Techniques Used: Expressing, Transfection, Western Blot, Plasmid Preparation, Cell Culture, Incubation

CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P
Figure Legend Snippet: CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P

Techniques Used: Incubation, Generated, Staining, Transfection, Pyrolysis Gas Chromatography, Western Blot

5) Product Images from "Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit"

Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit

Journal: PLoS ONE

doi: 10.1371/journal.pone.0202039

CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P
Figure Legend Snippet: CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P

Techniques Used: Expressing, Transfection, Western Blot, Plasmid Preparation, Cell Culture, Incubation

CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P
Figure Legend Snippet: CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P

Techniques Used: Incubation, Generated, Staining, Transfection, Pyrolysis Gas Chromatography, Western Blot

Related Articles

Incubation:

Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit
Article Snippet: .. Oxygen consumption analysis Astrocytes were subjected to Ru/R or CO/R for 20 h and further incubated with 0.5 μM antimycin A, 10 μM Nifedipine, 10 μM Compound C, or 2 mM EGTA for 4 h. Real time oxygen consumption in astrocytes was measured by the Oxygen Consumption Rate Assay Kit (Cayman, Ann Arbor, USA). ..

Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit
Article Snippet: .. Astrocytes were subjected to Ru/R or CO/R for 20 h and further incubated with 0.5 μM antimycin A, 10 μM Nifedipine, 10 μM Compound C, or 2 mM EGTA for 4 h. Real time oxygen consumption in astrocytes was measured by the Oxygen Consumption Rate Assay Kit (Cayman, Ann Arbor, USA). ..

other:

Article Title: Non-specific chemical inhibition of the Fanconi anemia pathway sensitizes cancer cells to cisplatin
Article Snippet: For subsequent studies, chemicals were purchased from Biomol (lactacystin, penitrem A, splitomicin, (±)-13-HODE), EMD biochemicals (actinomycin D, AG213, AG370, ALLN, alsterpaullone, α-amanitin, BAPTA-AM, bumetanide, CA-074-Me, curcumin, DRB, geldanamycin, Gö6976, HNMPA-(AM)3 , H-9, K-252c, MG132, nifedipine, propidium iodide, puromycin, roscovitine, SB218078, spermine NONOate, TPEN, trichostatin A, wortmannin), Cayman Chemical (leukotriene B3 ), Chembridge Corporation (5195243, 5315179, 5373662, 5656325, 5929407, 7012246), Fisher (chloroquine), Millenium Pharmaceutical (bortezomib), MP (3-methyladenine), Sigma (cisplatin, UCN-01), Tocris (ochratoxin A), VWR (17-AAG).

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    Cayman Chemical nifedipine
    The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM <t>nifedipine</t> (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p
    Nifedipine, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nifedipine/product/Cayman Chemical
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nifedipine - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

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    The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

    Journal: PLoS ONE

    Article Title: Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension

    doi: 10.1371/journal.pone.0195780

    Figure Lengend Snippet: The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

    Article Snippet: Pharmacological inhibitors nitrendipine, nifedipine, thapsigargin, U0126, mibefradil and Y-27632 were purchased from Cayman Chemicals (Ann Arbor, MI).

    Techniques: Standard Deviation, Incubation

    The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

    Journal: PLoS ONE

    Article Title: Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension

    doi: 10.1371/journal.pone.0195780

    Figure Lengend Snippet: The effect of calcium channel blockers on ET-1 and bradykinin induced cell contraction. Representative experiments showing bar graphs with standard deviation error bars for the effects of 10 uM nifedipine (Nif), 10 uM thapsigargin (thaps) (A, C), or mibefradil (Mib) (B, D) on ET-1 (A, B) or bradykinin (BK) (C, D) induced cell contraction. All inhibitors were pre-incubated with the cells for 1 h before addition of ET-1 or BK. #p

    Article Snippet: Pharmacological inhibitors nitrendipine, nifedipine, thapsigargin, U0126, mibefradil and Y-27632 were purchased from Cayman Chemicals (Ann Arbor, MI).

    Techniques: Standard Deviation, Incubation

    Larval zebrafish behavioral assays. (A) Doxycycline behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 7–8). In the first half‐hour behavioral assay (Test 1), larvae were treated with vehicle (−) or doxycycline (Dox). In the second half‐hour behavioral assay (Test 2), larvae were treated with vehicle, Dox, or PTZ . Movement greater than 20 mm/sec is shown in A’. (B) Metformin behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 30–66). In the first half‐hour behavioral assay, larvae were treated with vehicle or metformin (Met). In the second half‐hour behavioral assay, larvae were treated with vehicle, Met, or PTZ . Movement greater than 20 mm/sec is shown in B’. (C) Nifedipine behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38). In the first half‐hour behavioral assay, larvae were treated with vehicle or nifedipine (Nif). In the second half‐hour behavioral assay, larvae were treated with vehicle, Nif, or PTZ . Movement greater than 20 mm/sec is shown in C’. (D) Pyrantel tartrate behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38–40). In the first half‐hour behavioral assay, larvae were treated with vehicle or pyrantel tartrate (Pyr). In the second half‐hour behavioral assay, larvae were treated with vehicle, Pyr, or PTZ . Movement greater than 20 mm/sec is shown in D’. Bars represent mean ± SEM . **** P ‐value

    Journal: Annals of Clinical and Translational Neurology

    Article Title: Drug repositioning in epilepsy reveals novel antiseizure candidates

    doi: 10.1002/acn3.703

    Figure Lengend Snippet: Larval zebrafish behavioral assays. (A) Doxycycline behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 7–8). In the first half‐hour behavioral assay (Test 1), larvae were treated with vehicle (−) or doxycycline (Dox). In the second half‐hour behavioral assay (Test 2), larvae were treated with vehicle, Dox, or PTZ . Movement greater than 20 mm/sec is shown in A’. (B) Metformin behavioral assay. The average total distance moved at all speeds (mm) for each treatment group ( n = 30–66). In the first half‐hour behavioral assay, larvae were treated with vehicle or metformin (Met). In the second half‐hour behavioral assay, larvae were treated with vehicle, Met, or PTZ . Movement greater than 20 mm/sec is shown in B’. (C) Nifedipine behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38). In the first half‐hour behavioral assay, larvae were treated with vehicle or nifedipine (Nif). In the second half‐hour behavioral assay, larvae were treated with vehicle, Nif, or PTZ . Movement greater than 20 mm/sec is shown in C’. (D) Pyrantel tartrate behavioral assay. The average total distance moved at all speeds (mm) of each treatment group ( n = 38–40). In the first half‐hour behavioral assay, larvae were treated with vehicle or pyrantel tartrate (Pyr). In the second half‐hour behavioral assay, larvae were treated with vehicle, Pyr, or PTZ . Movement greater than 20 mm/sec is shown in D’. Bars represent mean ± SEM . **** P ‐value

    Article Snippet: Drug delivery Concentrated stock solutions for pentylenetetrazole (PTZ, Sigma), metformin (Cayman Chemical), doxycycline (Cayman Chemical), nifedipine (Cayman Chemical), and pyrantel tartrate (TargetMol) were made by dissolving each in either ddH2O (PTZ, doxycycline, metformin) or DMSO (nifedipine, pyrantel tartrate).

    Techniques: Behavioral Assay, Size-exclusion Chromatography

    CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P

    Journal: PLoS ONE

    Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit

    doi: 10.1371/journal.pone.0202039

    Figure Lengend Snippet: CO/R induces HIF-1α and ERRα expression via L-type Ca 2+ channels. ( A ) Astrocytes were transfected with 50 nM control or 50 nM HO-1 siRNA and subjected to Ru/R or CO/R. Indicated protein levels were analyzed using western blotting and quantified ( n = 4). ( B ) Astrocytes were treated with or without 10 μM Hemin for the time indicated ( n = 3). ( C ) Astrocytes were treated with 0, 5, or 10 μM Hemin for 8 h ( n = 3). ( D ) Cells were transfected with pcDNA3.1 mock or a pcDNA3.1/HO-1 vector and further cultured in fresh medium for 24 h ( n = 3). ( E ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO), 25 μM bilirubin (BR), and 25 μM FeCl (Fe 2+ ) in the presence or absence of 2 mM EGTA for 6 h. HIF-1α levels were analyzed using western blotting and quantified ( n = 5). ( F ) Astrocytes were pretreated with or without 25 μM SnPP, then incubated with 25 μM CORM-2 (CO) and 25 μM bilirubin (BR) in the presence or absence of 2 mM EGTA for 6 h ( n = 4). ( G ) Cells were subjected to Ru/R or CO/R, followed by treatment with 10 μM nifedipine (N), 0.3 μM ω-Conotoxin GVIA (C), 0.3 μM ω-Agatoxin TK (A), or 0.3 μM SNX 482 (S) for 4 h. HIF-1α and ERRα protein levels were quantified ( n = 7). ( H ) [Ca 2+ ] i was detected using the calcium-sensitive dye Fluo-4 AM ( n = 3). * P

    Article Snippet: Oxygen consumption analysis Astrocytes were subjected to Ru/R or CO/R for 20 h and further incubated with 0.5 μM antimycin A, 10 μM Nifedipine, 10 μM Compound C, or 2 mM EGTA for 4 h. Real time oxygen consumption in astrocytes was measured by the Oxygen Consumption Rate Assay Kit (Cayman, Ann Arbor, USA).

    Techniques: Expressing, Transfection, Western Blot, Plasmid Preparation, Cell Culture, Incubation

    CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P

    Journal: PLoS ONE

    Article Title: Heme oxygenase metabolites improve astrocytic mitochondrial function via a Ca2+-dependent HIF-1α/ERRα circuit

    doi: 10.1371/journal.pone.0202039

    Figure Lengend Snippet: CO/R enhances oxygen consumption. ( A-D ) Astrocytes were subjected to Ru/R or CO/R and further incubated with or without 0.5 μM Antimycin A (AMA), 2 mM EGTA, 10 μM Nifedipine, or 10 μM Compound C (Comp C) for 4 h. UT indicates untreated control cells. ( A-B ) Mitochondria oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit, and the quantified graph was generated at 45 min ( n = 5). ( C ) Mitochondrial oxygen consumption was detected by the Oxygen Consumption Rate Assay Kit ( n = 5). ( D ) Mitochondrial biogenesis was determined by staining with MitoTracker ( n = 5). ( E-F ) Astrocytes were transfected with 50 nM control, ERRα or PGC-1α siRNA, and subjected to Ru/R or CO/R. ( E ) Oxygen consumption was measured and quantified at 45 min ( n = 7). ( F ) Indicated proteins were detected via western blotting. * P

    Article Snippet: Oxygen consumption analysis Astrocytes were subjected to Ru/R or CO/R for 20 h and further incubated with 0.5 μM antimycin A, 10 μM Nifedipine, 10 μM Compound C, or 2 mM EGTA for 4 h. Real time oxygen consumption in astrocytes was measured by the Oxygen Consumption Rate Assay Kit (Cayman, Ann Arbor, USA).

    Techniques: Incubation, Generated, Staining, Transfection, Pyrolysis Gas Chromatography, Western Blot