phosphatase inhibitors  (Millipore)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Sodium orthovanadate
    Description:

    Catalog Number:
    S6508
    Price:
    None
    Applications:
    Sodium orthovandate was used to prepare the stop solution in dephosphorylation assay of insulin receptor kinase.26 It was one of the reagents used in the development of Matrix ChIP that utilizes surface-immobilized antibodies.27
    Buy from Supplier


    Structured Review

    Millipore phosphatase inhibitors
    Sodium orthovanadate

    https://www.bioz.com/result/phosphatase inhibitors/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphatase inhibitors - by Bioz Stars, 2021-06
    99/100 stars

    Images

    Related Articles

    Generated:

    Article Title: New Approaches to Prevent LEOPARD Syndrome-associated Cardiac Hypertrophy by Specifically Targeting Shp2-dependent Signaling *
    Article Snippet: Phenylephrine hydrochloride (PE) and U0126 were obtained from Sigma-Aldrich and dissolved in water or dimethyl sulfoxide, respectively. .. To induce strong tyrosine phosphorylation of focal adhesion kinase, NRCM were stimulated for 30 min with 10% fetal bovine serum in the presence of 0.1 m m pervanadate (generated by adding hydrogen peroxide to sodium orthovanadate stock solution (Sigma-Aldrich)). ..

    Incubation:

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation
    Article Snippet: The only exception was AD293vIII line, which was prepared by transfection of AD293 parental line with pIRESneo3-EGFRvIII using Fugene HD (Promega, Cat. no. E2311), selecting cells with neomycin treatment (500 µg/mL, InvivoGen, Cat. no. ANT-GN-1) and establishing monoclonal population, as described previously [ ]. .. Chemicals To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed. .. Following concentrations of inhibitors were used: DMSO (Sigma-Aldrich, Cat. no. D2438, solvent control), 10 μM erlotinib (Tarceva, Molecula; Cat. no. 89983631), 0.5 μM afatinib (Selleckchem, Cat. no. S1011); 5 μM gefitinib (CellSignaling; 4765), 5 μM lapatinib (Selleckchem, Cat. no. S1028), 300 nM flavopiridol (Selleckchem, Cat. no. S1230), 5 μM arry380 (Gentaur Europe, Cat. no. A8366), 10 μM Nu7441 (Selleckchem, Cat. no. S2638) and 10 μM ruxolitinib (Selleckchem, Cat. no. S1378).

    Article Title: Cyclic trans-phosphorylation in a homodimer as the predominant mechanism of EGFRvIII action and regulation
    Article Snippet: The only exception was AD293vIII line, which was prepared by transfection of AD293 parental line with pIRESneo3-EGFRvIII using Fugene HD (Promega, Cat. no. E2311), selecting cells with neomycin treatment (500 µg/mL, InvivoGen, Cat. no. ANT-GN-1) and establishing monoclonal population, as described previously [ ]. .. To analyse the effects of pharmacological inhibitors, cells were seeded in 6-well plates at 2.5x105 cells per well, incubated overnight to allow for adhesion and serum starved for subsequent 24 h. Serum free medium containing appropriate inhibitors was used to pre-treat cells for 30 min, unless indicated otherwise, prior to stimulation with 20 ng/mL EGF (Sigma-Aldrich, St Louis, MO, USA, Cat. no. E5036), 1 mM sodium orthovanadate – NaOVa (Na3 VO4 , Calbiochem, Cat. no. 567540), phenylarsine oxide – PAO (Sigma, Cat. no. P3075) or pervanadate – pV (obtained as described previously [ ]; using Catalase, Sigma, Cat. no. C1345 and H2 O2 Sigma, Cat. no. 31642) for 1 hour and lysed. .. Following concentrations of inhibitors were used: DMSO (Sigma-Aldrich, Cat. no. D2438, solvent control), 10 μM erlotinib (Tarceva, Molecula; Cat. no. 89983631), 0.5 μM afatinib (Selleckchem, Cat. no. S1011); 5 μM gefitinib (CellSignaling; 4765), 5 μM lapatinib (Selleckchem, Cat. no. S1028), 300 nM flavopiridol (Selleckchem, Cat. no. S1230), 5 μM arry380 (Gentaur Europe, Cat. no. A8366), 10 μM Nu7441 (Selleckchem, Cat. no. S2638) and 10 μM ruxolitinib (Selleckchem, Cat. no. S1378).

    Article Title: A single residue, arginine 65, is critical for the functional interaction of leukocyte associated inhibitory receptor (LAIR)-1 with collagens 1
    Article Snippet: After 3 h of incubation at 37 °C cells are harvested and conjugate formation was analyzed in a FACScalibur cytometer (BD Biosciences). .. LAIR-1 wt or LAIR-1 R65K transfected K562 cells were treated with collagen type I at 1 μg/107 cells/ml in medium without serum at 37 °C for 20 min. For pervanadate treatment, cells were incubated for 20 min with freshly prepared sodium pervanadate (0.1 mM sodium orthovanadate and 10 mM hydrogen peroxide from Sigma-Aldrich) in medium without serum at 37 °C. ..

    Positive Control:

    Article Title: Activation of Triggering Receptor Expressed on Myeloid Cells-1 on Human Neutrophils by Marburg and Ebola Viruses
    Article Snippet: Briefly, human neutrophils (107 /100 μl of HBSS with Ca2+ /Mg2+ ) were incubated with inactivated MARVCi67 (iMARVCi67 ) (1 μg/100 μl) or iEBOVZaire (1 μg/100 μl) for 0, 1, 5, or 10 min at 37°C. .. Pervanadate (sodium orthovanadate plus H2 O2 ; Sigma, St. Louis, MO) was used as a positive control ( ). .. After stimulation, neutrophils were lysed and lysates were immune precipitated using polyclonal anti-DAP12 antibody (kindly provided by D. McVicar, NCI, Frederick, MD), separated on a 10% Bis-Tris NuPAGE gel (Invitrogen, Carlsbad, CA), and transferred onto activated polyvinylidene difluoride membranes (Millipore, Billerica, MA).

    Article Title: Aberrant STAT5 and PI3K/mTOR pathway signaling occurs in human CRLF2-rearranged B-precursor acute lymphoblastic leukemia
    Article Snippet: .. Pervanadate (125μM; prepared from sodium orthovanadate and 3% hydrogen peroxide; Sigma-Aldrich), an irreversible protein tyrosine phosphatase inhibitor, was also used as a positive control to elicit maximal phosphorylation of each signaling protein. .. Cells were immediately lysed in NP-40 cell lysis buffer (Invitrogen) supplemented with 1% protease and phosphatase inhibitors (Calbiochem).

    Transfection:

    Article Title: A single residue, arginine 65, is critical for the functional interaction of leukocyte associated inhibitory receptor (LAIR)-1 with collagens 1
    Article Snippet: After 3 h of incubation at 37 °C cells are harvested and conjugate formation was analyzed in a FACScalibur cytometer (BD Biosciences). .. LAIR-1 wt or LAIR-1 R65K transfected K562 cells were treated with collagen type I at 1 μg/107 cells/ml in medium without serum at 37 °C for 20 min. For pervanadate treatment, cells were incubated for 20 min with freshly prepared sodium pervanadate (0.1 mM sodium orthovanadate and 10 mM hydrogen peroxide from Sigma-Aldrich) in medium without serum at 37 °C. ..

    Concentration Assay:

    Article Title: Adaptive potentiation in rod photoreceptors after light exposure
    Article Snippet: Final concentrations were chosen based on literature reports of the EC50 for each compound: Tyr538 at 1 µM lacks specificity for inhibiting the IGF1R and the IR ( ); Tyr1024 at 250 nM is significantly more specific for IGF-1R than the IR ( ); HNMPA(AM)3 at 200 µM is specific for IR kinase inhibition ( ). .. The tyrosine phosphatase inhibitor sodium orthovanadate (Na3 VO4 ; Sigma-Aldrich) stock was prepared in diH2 0 and adjusted to pH 10.0 before being added to Locke’s buffer for a final experimental solution concentration of 200 µM and pH of 7.4. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Millipore faah inhibitor
    Effects of the <t>FAAH</t> inhibitor <t>URB597</t> (30 ng per 0.5 μl) and the CB1 receptor antagonist AM251 (300 ng per 0.5 μl) on CS-fear memory reconsolidation. Administration of URB597 immediately after the reactivation session produced a small, transient enhancement of CS-fear memory reconsolidation at 24 h, but not 8 days, after reactivation. AM251 persistently impaired memory reconsolidation when compared with vehicle and URB597-treated rats after both 24 h and 8 days after the reactivation session. Data are presented as means±SEM. Group sizes were VEH, n =10; URB597, n =12; and AM251, n =10.
    Faah Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/faah inhibitor/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    faah inhibitor - by Bioz Stars, 2021-06
    97/100 stars
      Buy from Supplier

    99
    Millipore tyrosine phosphatase inhibitor sodium orthovanadate
    Kinase and <t>phosphatase</t> inhibitors influence light-induced potentiation in isolated retina recordings. All figures show a dark-adapted response (black trace, average of three to five responses), a response recorded 3–5 s after 3-min saturating light (red trace, single response), and a third response, recorded 20–30 s later, representing recovery (blue trace, single response). (A–D) Each panel shows two potentiation experiments on a single retina before (left) and after (right) the application of the indicated drug or drugs. (A) Potentiation is present in control solution (left), showing a 46% increase in peak amplitude before application of HNMPA(AM) 3 , a specific blocker of IR kinase activity. The presence of 200 µM HNMPA(AM) 3 did not affect the potentiation (right), as the potentiation (red trace) persists in the presence of the <t>inhibitor.</t> (B) Potentiation is present in control solution (left), showing a 36% increase in amplitude before application of Tyr1024, a specific blocker of IGF-1R kinase activity. The presence of 250 nM Tyr1024 eliminated the potentiation (right) after a conditioning light; in fact, there was a decrease in amplitude that recovered with time (blue trace). (C) Application of both 250 nM Tyr1024 and 200 µM HNMPA(AM) 3 eliminates potentiation and the amplitude reduction after light exposure seen with Try1024 alone. (D) Potentiation is present in control solution (left), showing a 44% increase in amplitude before application of <t>orthovanadate,</t> a broad-acting inhibitor of <t>tyrosine</t> phosphatases. The presence of 200 µM orthovanadate eliminates the potentiation (right).
    Tyrosine Phosphatase Inhibitor Sodium Orthovanadate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tyrosine phosphatase inhibitor sodium orthovanadate/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tyrosine phosphatase inhibitor sodium orthovanadate - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    94
    Millipore hck inhibitor pp1
    DON-induced rRNA cleavage in RAW 264.7 involves PKR, <t>Hck,</t> p38, p53, and caspases. Cells were preincubated with (A) C-16 (0.1 or 0.3 μM), (B) <t>PP1</t> (5 or 25 μM), (C) SB 203580 (1 or 5 μM), (D) pifithrin-α (80 or 100 μM), (E) pifithrin-μ (10 or 25 μM), or (F) Z-VAD-FMK (50 or 100 μM) for 1 h and then with DON (1000 ng/ml) for 6 h. RNAs were purified and analyzed by capillary electrophoresis. Results are representative of three separate experiments. Arrows indicate that fragmentation to major cleavage peaks is suppressed by inhibitor.
    Hck Inhibitor Pp1, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hck inhibitor pp1/product/Millipore
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hck inhibitor pp1 - by Bioz Stars, 2021-06
    94/100 stars
      Buy from Supplier

    92
    Millipore anti neurabin
    Neonatal Dnmt inhibition increases dendritic spine density on POA neurons and masculinizes behavior (a) Animals were treated with the Dnmt inhibitors Zeb or RG108 on PN0 and PN1. As adults, animals were gonadectomized and implanted with testosterone-releasing capsules to achieve stable adult male circulating testosterone levels. Male sexual behavior was assessed 20 days later and brains were collected at the completion of behavioral testing. ( b ) Golgi-Cox impregnated adult POA neurons were quantified for dendritic spine density. Neonatal Dnmt inhibition by either Zeb or RG108 in females masculinized dendritic spine density of adults (ANOVA, F(3,22) = 4.92, p = 0.0091). Scale bar = 10 µm (c ) Zeb treatment on PN0 PN1 significantly increased <t>Neurabin</t> II protein levels in the adult female POA (F(3,32) = 4.69, p = 0.0079). Cropped representative western bands are shown and full length blots are provided in Supplementary Figure 10 . ( d ) Neonatal Zeb treatment increased mounts (F(3,25) = 22.18, p
    Anti Neurabin, supplied by Millipore, 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/anti neurabin/product/Millipore
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti neurabin - by Bioz Stars, 2021-06
    92/100 stars
      Buy from Supplier

    Image Search Results


    Effects of the FAAH inhibitor URB597 (30 ng per 0.5 μl) and the CB1 receptor antagonist AM251 (300 ng per 0.5 μl) on CS-fear memory reconsolidation. Administration of URB597 immediately after the reactivation session produced a small, transient enhancement of CS-fear memory reconsolidation at 24 h, but not 8 days, after reactivation. AM251 persistently impaired memory reconsolidation when compared with vehicle and URB597-treated rats after both 24 h and 8 days after the reactivation session. Data are presented as means±SEM. Group sizes were VEH, n =10; URB597, n =12; and AM251, n =10.

    Journal: Neuropsychopharmacology

    Article Title: The CB1 receptor antagonist AM251 impairs reconsolidation of pavlovian fear memory in the rat basolateral amygdala

    doi: 10.1038/npp.2014.103

    Figure Lengend Snippet: Effects of the FAAH inhibitor URB597 (30 ng per 0.5 μl) and the CB1 receptor antagonist AM251 (300 ng per 0.5 μl) on CS-fear memory reconsolidation. Administration of URB597 immediately after the reactivation session produced a small, transient enhancement of CS-fear memory reconsolidation at 24 h, but not 8 days, after reactivation. AM251 persistently impaired memory reconsolidation when compared with vehicle and URB597-treated rats after both 24 h and 8 days after the reactivation session. Data are presented as means±SEM. Group sizes were VEH, n =10; URB597, n =12; and AM251, n =10.

    Article Snippet: URB597 , a FAAH inhibitor (URB (cyclohexylcarbamic acid 3 ́ -carbamoyl-biphenyl-3-yl ester), 30 ng per 0.5 μl per side, Sigma-Aldrich, Dorset, UK), the CB1R antagonist AM251 ( ; AM ( N -(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide), 300 ng per 0.5 μl per side, Tocris, Bristol, UK), and GABAA receptor antagonist 1(S),9(R)-(−)-Bicuculline methiodide (BIC; 50 ng per 0.5 μl per side, Sigma-Aldrich) were dissolved in a vehicle (VEH) containing 5% polyethylene glycol, 5% Tween-80, and 90% saline.

    Techniques: Produced

    Effects of the FAAH inhibitor URB597 (30 ng per 0.5 μl) and the CB1 receptor antagonist AM251 (300 ng per 0.5 μl) on CS-fear memory reconsolidation in rats not exposed to the memory reactivation session. Administration of URB597 or AM251 in the absence of memory reactivation had no effect on the retrieval of the CS-fear memory both 24 h and 8 days after administration. Data are presented as means±SEM. Group sizes were VEH, n =8; URB597, n =8; and AM251, n =8.

    Journal: Neuropsychopharmacology

    Article Title: The CB1 receptor antagonist AM251 impairs reconsolidation of pavlovian fear memory in the rat basolateral amygdala

    doi: 10.1038/npp.2014.103

    Figure Lengend Snippet: Effects of the FAAH inhibitor URB597 (30 ng per 0.5 μl) and the CB1 receptor antagonist AM251 (300 ng per 0.5 μl) on CS-fear memory reconsolidation in rats not exposed to the memory reactivation session. Administration of URB597 or AM251 in the absence of memory reactivation had no effect on the retrieval of the CS-fear memory both 24 h and 8 days after administration. Data are presented as means±SEM. Group sizes were VEH, n =8; URB597, n =8; and AM251, n =8.

    Article Snippet: URB597 , a FAAH inhibitor (URB (cyclohexylcarbamic acid 3 ́ -carbamoyl-biphenyl-3-yl ester), 30 ng per 0.5 μl per side, Sigma-Aldrich, Dorset, UK), the CB1R antagonist AM251 ( ; AM ( N -(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide), 300 ng per 0.5 μl per side, Tocris, Bristol, UK), and GABAA receptor antagonist 1(S),9(R)-(−)-Bicuculline methiodide (BIC; 50 ng per 0.5 μl per side, Sigma-Aldrich) were dissolved in a vehicle (VEH) containing 5% polyethylene glycol, 5% Tween-80, and 90% saline.

    Techniques:

    Effects of the FAAH inhibitor URB597 and the CB1 receptor antagonist AM251 on CS-fear memory reconsolidation. Administration of URB597 or AM251 before memory reactivation had no effect on the retrieval of the CS-fear memory at reactivation and did not alter expression of freezing response at tests conducted 24 h or 8 days later. Group sizes were VEH, n =9; URB597, n =10; and AM251, n =9.

    Journal: Neuropsychopharmacology

    Article Title: The CB1 receptor antagonist AM251 impairs reconsolidation of pavlovian fear memory in the rat basolateral amygdala

    doi: 10.1038/npp.2014.103

    Figure Lengend Snippet: Effects of the FAAH inhibitor URB597 and the CB1 receptor antagonist AM251 on CS-fear memory reconsolidation. Administration of URB597 or AM251 before memory reactivation had no effect on the retrieval of the CS-fear memory at reactivation and did not alter expression of freezing response at tests conducted 24 h or 8 days later. Group sizes were VEH, n =9; URB597, n =10; and AM251, n =9.

    Article Snippet: URB597 , a FAAH inhibitor (URB (cyclohexylcarbamic acid 3 ́ -carbamoyl-biphenyl-3-yl ester), 30 ng per 0.5 μl per side, Sigma-Aldrich, Dorset, UK), the CB1R antagonist AM251 ( ; AM ( N -(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamide), 300 ng per 0.5 μl per side, Tocris, Bristol, UK), and GABAA receptor antagonist 1(S),9(R)-(−)-Bicuculline methiodide (BIC; 50 ng per 0.5 μl per side, Sigma-Aldrich) were dissolved in a vehicle (VEH) containing 5% polyethylene glycol, 5% Tween-80, and 90% saline.

    Techniques: Expressing

    Kinase and phosphatase inhibitors influence light-induced potentiation in isolated retina recordings. All figures show a dark-adapted response (black trace, average of three to five responses), a response recorded 3–5 s after 3-min saturating light (red trace, single response), and a third response, recorded 20–30 s later, representing recovery (blue trace, single response). (A–D) Each panel shows two potentiation experiments on a single retina before (left) and after (right) the application of the indicated drug or drugs. (A) Potentiation is present in control solution (left), showing a 46% increase in peak amplitude before application of HNMPA(AM) 3 , a specific blocker of IR kinase activity. The presence of 200 µM HNMPA(AM) 3 did not affect the potentiation (right), as the potentiation (red trace) persists in the presence of the inhibitor. (B) Potentiation is present in control solution (left), showing a 36% increase in amplitude before application of Tyr1024, a specific blocker of IGF-1R kinase activity. The presence of 250 nM Tyr1024 eliminated the potentiation (right) after a conditioning light; in fact, there was a decrease in amplitude that recovered with time (blue trace). (C) Application of both 250 nM Tyr1024 and 200 µM HNMPA(AM) 3 eliminates potentiation and the amplitude reduction after light exposure seen with Try1024 alone. (D) Potentiation is present in control solution (left), showing a 44% increase in amplitude before application of orthovanadate, a broad-acting inhibitor of tyrosine phosphatases. The presence of 200 µM orthovanadate eliminates the potentiation (right).

    Journal: The Journal of General Physiology

    Article Title: Adaptive potentiation in rod photoreceptors after light exposure

    doi: 10.1085/jgp.201411163

    Figure Lengend Snippet: Kinase and phosphatase inhibitors influence light-induced potentiation in isolated retina recordings. All figures show a dark-adapted response (black trace, average of three to five responses), a response recorded 3–5 s after 3-min saturating light (red trace, single response), and a third response, recorded 20–30 s later, representing recovery (blue trace, single response). (A–D) Each panel shows two potentiation experiments on a single retina before (left) and after (right) the application of the indicated drug or drugs. (A) Potentiation is present in control solution (left), showing a 46% increase in peak amplitude before application of HNMPA(AM) 3 , a specific blocker of IR kinase activity. The presence of 200 µM HNMPA(AM) 3 did not affect the potentiation (right), as the potentiation (red trace) persists in the presence of the inhibitor. (B) Potentiation is present in control solution (left), showing a 36% increase in amplitude before application of Tyr1024, a specific blocker of IGF-1R kinase activity. The presence of 250 nM Tyr1024 eliminated the potentiation (right) after a conditioning light; in fact, there was a decrease in amplitude that recovered with time (blue trace). (C) Application of both 250 nM Tyr1024 and 200 µM HNMPA(AM) 3 eliminates potentiation and the amplitude reduction after light exposure seen with Try1024 alone. (D) Potentiation is present in control solution (left), showing a 44% increase in amplitude before application of orthovanadate, a broad-acting inhibitor of tyrosine phosphatases. The presence of 200 µM orthovanadate eliminates the potentiation (right).

    Article Snippet: The tyrosine phosphatase inhibitor sodium orthovanadate (Na3 VO4 ; Sigma-Aldrich) stock was prepared in diH2 0 and adjusted to pH 10.0 before being added to Locke’s buffer for a final experimental solution concentration of 200 µM and pH of 7.4.

    Techniques: Isolation, Activity Assay

    DON-induced rRNA cleavage in RAW 264.7 involves PKR, Hck, p38, p53, and caspases. Cells were preincubated with (A) C-16 (0.1 or 0.3 μM), (B) PP1 (5 or 25 μM), (C) SB 203580 (1 or 5 μM), (D) pifithrin-α (80 or 100 μM), (E) pifithrin-μ (10 or 25 μM), or (F) Z-VAD-FMK (50 or 100 μM) for 1 h and then with DON (1000 ng/ml) for 6 h. RNAs were purified and analyzed by capillary electrophoresis. Results are representative of three separate experiments. Arrows indicate that fragmentation to major cleavage peaks is suppressed by inhibitor.

    Journal: Toxicological Sciences

    Article Title: Targets and Intracellular Signaling Mechanisms for Deoxynivalenol-Induced Ribosomal RNA Cleavage

    doi: 10.1093/toxsci/kfs134

    Figure Lengend Snippet: DON-induced rRNA cleavage in RAW 264.7 involves PKR, Hck, p38, p53, and caspases. Cells were preincubated with (A) C-16 (0.1 or 0.3 μM), (B) PP1 (5 or 25 μM), (C) SB 203580 (1 or 5 μM), (D) pifithrin-α (80 or 100 μM), (E) pifithrin-μ (10 or 25 μM), or (F) Z-VAD-FMK (50 or 100 μM) for 1 h and then with DON (1000 ng/ml) for 6 h. RNAs were purified and analyzed by capillary electrophoresis. Results are representative of three separate experiments. Arrows indicate that fragmentation to major cleavage peaks is suppressed by inhibitor.

    Article Snippet: DON, anisomycin, PKR inhibitor C-16, and Hck inhibitor PP1 were purchased from Sigma-Aldrich (St Louis, MO).

    Techniques: Purification, Electrophoresis

    Neonatal Dnmt inhibition increases dendritic spine density on POA neurons and masculinizes behavior (a) Animals were treated with the Dnmt inhibitors Zeb or RG108 on PN0 and PN1. As adults, animals were gonadectomized and implanted with testosterone-releasing capsules to achieve stable adult male circulating testosterone levels. Male sexual behavior was assessed 20 days later and brains were collected at the completion of behavioral testing. ( b ) Golgi-Cox impregnated adult POA neurons were quantified for dendritic spine density. Neonatal Dnmt inhibition by either Zeb or RG108 in females masculinized dendritic spine density of adults (ANOVA, F(3,22) = 4.92, p = 0.0091). Scale bar = 10 µm (c ) Zeb treatment on PN0 PN1 significantly increased Neurabin II protein levels in the adult female POA (F(3,32) = 4.69, p = 0.0079). Cropped representative western bands are shown and full length blots are provided in Supplementary Figure 10 . ( d ) Neonatal Zeb treatment increased mounts (F(3,25) = 22.18, p

    Journal: Nature neuroscience

    Article Title: Brain feminization requires active repression of masculinization via DNA methylation

    doi: 10.1038/nn.3988

    Figure Lengend Snippet: Neonatal Dnmt inhibition increases dendritic spine density on POA neurons and masculinizes behavior (a) Animals were treated with the Dnmt inhibitors Zeb or RG108 on PN0 and PN1. As adults, animals were gonadectomized and implanted with testosterone-releasing capsules to achieve stable adult male circulating testosterone levels. Male sexual behavior was assessed 20 days later and brains were collected at the completion of behavioral testing. ( b ) Golgi-Cox impregnated adult POA neurons were quantified for dendritic spine density. Neonatal Dnmt inhibition by either Zeb or RG108 in females masculinized dendritic spine density of adults (ANOVA, F(3,22) = 4.92, p = 0.0091). Scale bar = 10 µm (c ) Zeb treatment on PN0 PN1 significantly increased Neurabin II protein levels in the adult female POA (F(3,32) = 4.69, p = 0.0079). Cropped representative western bands are shown and full length blots are provided in Supplementary Figure 10 . ( d ) Neonatal Zeb treatment increased mounts (F(3,25) = 22.18, p

    Article Snippet: Membranes were blocked in 5% nonfat milk in 0.1% Tween TBS (T-TTBS) for 1 hr at room temperature and then incubated overnight at 4°C in either anti-Dnmt1 (Santa Cruz, Cat.# SC–20701; 1:2000; 1DegreeBio ID: 1DB-001-0001405395), anti-Dnmt3a (Cell Signaling, Cat.# 2160; 1:1000; 1DegreeBio ID: 1DB-001-0000807788), anti-Dnmt3b (Cell Signaling, Cat.# 2161; 1:1500; 1DegreeBio ID: 1DB-001-0000807789), or anti-Neurabin (Millipore, Cat.# 06–852;1:000; 1DegreeBio ID: 1DB-001-0000850617) in 2.5 % milk in TTBS, followed by a 30-minute incubation in goat anti-rabbit HRP conjugated IgG (Cell Signaling; 1:3000) in TTBS.

    Techniques: Inhibition, Western Blot

    Dnmt inhibition outside of the critical period increases dendritic spine markers in the POA and masculinizes behavior (a) Animals were treated with Zeb or estradiol at PN10 and PN11. In adulthood, animals were gonadectomized and implanted with testosterone-releasing capsules and male sexual behavior was assessed 20 days later. Brains were collected and Neurabin II levels were quantified in the POA following the completion of three weeks of behavioral testing. ( b ) Zeb administration after the close of the critical period for sexual differentiation of the brain masculinized Neurabin II protein levels ( a priori t-test between females and females + zeb: t(8) = –2.833, p = 0.0221), although there was no main effect across all treatment groups (ANOVA, F(4,22) = 2.22, p = 0.1002). Cropped representative western bands are shown and full length blots are provided in Supplementary Figure 10 . ( c ) Number of mounts (F(4,27) = 5.23, p = 0.003), the latency to mount (F(4,26) = 2.44, p = 0.0718, a priori t-test between females and females + zeb: t(10) = 2.862, p = 0.0169), and thrust latency (F(4,21) = 6.67, p = 0.0013) were also masculinized by Dnmt inhibition whereas estradiol treatment administered at this same time point did not induce masculinization of brain or behavior. Although there were large differences in the number of thrusts between males and females (F(4,27) = 14.861, p

    Journal: Nature neuroscience

    Article Title: Brain feminization requires active repression of masculinization via DNA methylation

    doi: 10.1038/nn.3988

    Figure Lengend Snippet: Dnmt inhibition outside of the critical period increases dendritic spine markers in the POA and masculinizes behavior (a) Animals were treated with Zeb or estradiol at PN10 and PN11. In adulthood, animals were gonadectomized and implanted with testosterone-releasing capsules and male sexual behavior was assessed 20 days later. Brains were collected and Neurabin II levels were quantified in the POA following the completion of three weeks of behavioral testing. ( b ) Zeb administration after the close of the critical period for sexual differentiation of the brain masculinized Neurabin II protein levels ( a priori t-test between females and females + zeb: t(8) = –2.833, p = 0.0221), although there was no main effect across all treatment groups (ANOVA, F(4,22) = 2.22, p = 0.1002). Cropped representative western bands are shown and full length blots are provided in Supplementary Figure 10 . ( c ) Number of mounts (F(4,27) = 5.23, p = 0.003), the latency to mount (F(4,26) = 2.44, p = 0.0718, a priori t-test between females and females + zeb: t(10) = 2.862, p = 0.0169), and thrust latency (F(4,21) = 6.67, p = 0.0013) were also masculinized by Dnmt inhibition whereas estradiol treatment administered at this same time point did not induce masculinization of brain or behavior. Although there were large differences in the number of thrusts between males and females (F(4,27) = 14.861, p

    Article Snippet: Membranes were blocked in 5% nonfat milk in 0.1% Tween TBS (T-TTBS) for 1 hr at room temperature and then incubated overnight at 4°C in either anti-Dnmt1 (Santa Cruz, Cat.# SC–20701; 1:2000; 1DegreeBio ID: 1DB-001-0001405395), anti-Dnmt3a (Cell Signaling, Cat.# 2160; 1:1000; 1DegreeBio ID: 1DB-001-0000807788), anti-Dnmt3b (Cell Signaling, Cat.# 2161; 1:1500; 1DegreeBio ID: 1DB-001-0000807789), or anti-Neurabin (Millipore, Cat.# 06–852;1:000; 1DegreeBio ID: 1DB-001-0000850617) in 2.5 % milk in TTBS, followed by a 30-minute incubation in goat anti-rabbit HRP conjugated IgG (Cell Signaling; 1:3000) in TTBS.

    Techniques: Inhibition, Western Blot