fumonisin b1  (Millipore)


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    Name:
    Fumonisin B1
    Description:
    Fumonisins are a family of mycotoxins that are characterized by their structurally related homologs They are generally produced by fungi of Fusarium species
    Catalog Number:
    32936
    Price:
    None
    Applications:
    Refer to the product's Certificate of Analysis for more information on a suitable instrument technique. Contact Technical Service for further support.
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    Structured Review

    Millipore fumonisin b1
    Fumonisin B1
    Fumonisins are a family of mycotoxins that are characterized by their structurally related homologs They are generally produced by fungi of Fusarium species
    https://www.bioz.com/result/fumonisin b1/product/Millipore
    Average 96 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    fumonisin b1 - by Bioz Stars, 2020-09
    96/100 stars

    Images

    1) Product Images from "Two New Fluorogenic Aptasensors Based on Capped Mesoporous Silica Nanoparticles to Detect Ochratoxin A"

    Article Title: Two New Fluorogenic Aptasensors Based on Capped Mesoporous Silica Nanoparticles to Detect Ochratoxin A

    Journal: ChemistryOpen

    doi: 10.1002/open.201700106

    Release of rhodamine B from solid S3 (in black) and S5 (in grey) in the presence of OTA, aflatoxin B1, and fumonisin B1.
    Figure Legend Snippet: Release of rhodamine B from solid S3 (in black) and S5 (in grey) in the presence of OTA, aflatoxin B1, and fumonisin B1.

    Techniques Used:

    2) Product Images from "Cytokine secretion requires phosphatidylcholine synthesis"

    Article Title: Cytokine secretion requires phosphatidylcholine synthesis

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200706152

    Response of TNFα secretion to different treatments. (A) Wild-type (WT; white bars) or CCTα-deficient (KO; black bars) macrophages were treated with 10 ng/ml LPS for 18 h. At the same time, 100 μM LysoPC, 15 μM edelfosine, or solvent vehicle (0.1% ethanol) were added to the medium as indicated. The amount of TNFα secreted into the culture medium was determined using a Quantikine kit, and results are the mean values normalized to cellular protein ( n = 8) obtained from four mice of each genotype. (B) Wild-type cells were treated with 10 ng/ml LPS alone, with LPS plus 10 μM fumonisin B1 to inhibit ceramide synthesis, with LPS plus 50 μM propranolol to inhibit the PtdOH P'tse, with LPS plus 15 mM 1-butanol to inhibit the phospholipase D, and with LPS plus 20 mM 2-butanol as an alcohol control for 8 h. Data are the mean of determinations from four individual mice of each genotype. Error bars indicate mean ± SE. *, P
    Figure Legend Snippet: Response of TNFα secretion to different treatments. (A) Wild-type (WT; white bars) or CCTα-deficient (KO; black bars) macrophages were treated with 10 ng/ml LPS for 18 h. At the same time, 100 μM LysoPC, 15 μM edelfosine, or solvent vehicle (0.1% ethanol) were added to the medium as indicated. The amount of TNFα secreted into the culture medium was determined using a Quantikine kit, and results are the mean values normalized to cellular protein ( n = 8) obtained from four mice of each genotype. (B) Wild-type cells were treated with 10 ng/ml LPS alone, with LPS plus 10 μM fumonisin B1 to inhibit ceramide synthesis, with LPS plus 50 μM propranolol to inhibit the PtdOH P'tse, with LPS plus 15 mM 1-butanol to inhibit the phospholipase D, and with LPS plus 20 mM 2-butanol as an alcohol control for 8 h. Data are the mean of determinations from four individual mice of each genotype. Error bars indicate mean ± SE. *, P

    Techniques Used: Mouse Assay

    3) Product Images from "Hepatic triglyceride accumulation via endoplasmic reticulum stress-induced SREBP-1 activation is regulated by ceramide synthases"

    Article Title: Hepatic triglyceride accumulation via endoplasmic reticulum stress-induced SREBP-1 activation is regulated by ceramide synthases

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/s12276-019-0340-1

    Bortezomib-induced increase in CerS2 alleviates palmitate-induced ER stress. The cells were pretreated with bortezomib (BT, 50 nM) and fumonisin B1 (FB1, 20 µM) 12 h before palmitate (500 μM) treatment. After 12 h of palmitate treatment, CerS, ER stress markers, SREBP-1, and INSIG-1 protein levels were examined using Western blotting. The images are representative images of three independent experiments. CerS ceramide synthase, Con control, BT Bortezomib, eIF2α eukaryotic initiation factor 2α, INSIG-1 Insulin-induced gene 1 protein, m-SREBP-1 mature form of sterol regulatory element-binding protein 1, p-SREBP-1 precursor form of sterol regulatory element-binding protein 1, PERK Protein kinase RNA-like endoplasmic reticulum kinase
    Figure Legend Snippet: Bortezomib-induced increase in CerS2 alleviates palmitate-induced ER stress. The cells were pretreated with bortezomib (BT, 50 nM) and fumonisin B1 (FB1, 20 µM) 12 h before palmitate (500 μM) treatment. After 12 h of palmitate treatment, CerS, ER stress markers, SREBP-1, and INSIG-1 protein levels were examined using Western blotting. The images are representative images of three independent experiments. CerS ceramide synthase, Con control, BT Bortezomib, eIF2α eukaryotic initiation factor 2α, INSIG-1 Insulin-induced gene 1 protein, m-SREBP-1 mature form of sterol regulatory element-binding protein 1, p-SREBP-1 precursor form of sterol regulatory element-binding protein 1, PERK Protein kinase RNA-like endoplasmic reticulum kinase

    Techniques Used: Western Blot, Binding Assay

    The effects of ceramide on palmitate-induced ER stress are different in Hep3B cells depending on ceramide acyl chain length. a Representative Western blots of ER stress markers upon treatment with palmitate (500 μM) and ceramides (1 μM) with various acyl chain lengths. b Representative Western blots of ER stress markers upon treatment with palmitate in CerS5- or CerS6-overexpressing cells. c Representative Western blots of ER stress markers upon treatment with palmitate in CerS2-overexpressing cells. For CerS inhibition, the cells were pretreated with fumonisin b1 (FB1, 20 µM) 1 h before palmitate treatment. Three independent experiments are presented. CerS ceramide synthase, CHOP CCAAT-enhancer-binding protein homologous protein, Con control, eIF2α eukaryotic initiation factor 2α, GRP78 , 78 kDa glucose-regulated protein, PERK Protein kinase RNA-like endoplasmic reticulum kinase
    Figure Legend Snippet: The effects of ceramide on palmitate-induced ER stress are different in Hep3B cells depending on ceramide acyl chain length. a Representative Western blots of ER stress markers upon treatment with palmitate (500 μM) and ceramides (1 μM) with various acyl chain lengths. b Representative Western blots of ER stress markers upon treatment with palmitate in CerS5- or CerS6-overexpressing cells. c Representative Western blots of ER stress markers upon treatment with palmitate in CerS2-overexpressing cells. For CerS inhibition, the cells were pretreated with fumonisin b1 (FB1, 20 µM) 1 h before palmitate treatment. Three independent experiments are presented. CerS ceramide synthase, CHOP CCAAT-enhancer-binding protein homologous protein, Con control, eIF2α eukaryotic initiation factor 2α, GRP78 , 78 kDa glucose-regulated protein, PERK Protein kinase RNA-like endoplasmic reticulum kinase

    Techniques Used: Western Blot, Inhibition, Binding Assay

    4) Product Images from "Deguelin Induces Both Apoptosis and Autophagy in Cultured Head and Neck Squamous Cell Carcinoma Cells"

    Article Title: Deguelin Induces Both Apoptosis and Autophagy in Cultured Head and Neck Squamous Cell Carcinoma Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0054736

    Deguelin activates AMPK signaling. Hep-2 cells were exposed to 100 µM of deguelin for indicated time points, followed by western blot detecting phospho- and total- level of LKB1-AMPK-ACC using specific antibodies (A). Hep-2 cells were transfected with scramble or AMPKα1RNAi (100 nM each) for 48 hours. Western blot was utilized to test AMPKα1 expression after transfection. Successfully AMPK knockdown cells and their parental cells were treated with deguelin (100 µM) in the presence or absence of AICAR (1 mM), phospho- and total- level of AMPKα1 were analyzed in (B–C), MTT assay was then used to test cell viability after 48 hours (D). Hep-2 cells were pretreated with fumonisin B1 (20 µM), C6-ceramide (10 µg/ml) or NAC (500 µM) for 1 hr before deguelin exposure for indicated time points, phospho- and total level of AMPKα1 as well as tubulin were tested by western blots (E–F). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.
    Figure Legend Snippet: Deguelin activates AMPK signaling. Hep-2 cells were exposed to 100 µM of deguelin for indicated time points, followed by western blot detecting phospho- and total- level of LKB1-AMPK-ACC using specific antibodies (A). Hep-2 cells were transfected with scramble or AMPKα1RNAi (100 nM each) for 48 hours. Western blot was utilized to test AMPKα1 expression after transfection. Successfully AMPK knockdown cells and their parental cells were treated with deguelin (100 µM) in the presence or absence of AICAR (1 mM), phospho- and total- level of AMPKα1 were analyzed in (B–C), MTT assay was then used to test cell viability after 48 hours (D). Hep-2 cells were pretreated with fumonisin B1 (20 µM), C6-ceramide (10 µg/ml) or NAC (500 µM) for 1 hr before deguelin exposure for indicated time points, phospho- and total level of AMPKα1 as well as tubulin were tested by western blots (E–F). The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.

    Techniques Used: Western Blot, Transfection, Expressing, MTT Assay

    Deguelin induces cellular ceramide synthesis. Hep-2 cells were exposed to 100 µM of deguelin with or without fumonisin B1 (20 µM) for indicated time points, intracellular ceramide level was analyzed using methods mentioned above (A). Hep-2 cells were treated with fumonisin B1 (20 µM) or C6-ceramide (10 µg/ml), followed by deguelin (100 µM) exposure, MTT assay was used to test cell viability after 72 hours (B), Hoechst staining (C) and enzyme-linked immunosorbent cell apoptosis assay (D) were utilized to test Hep-2 cell apoptosis after 48 hours. The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.
    Figure Legend Snippet: Deguelin induces cellular ceramide synthesis. Hep-2 cells were exposed to 100 µM of deguelin with or without fumonisin B1 (20 µM) for indicated time points, intracellular ceramide level was analyzed using methods mentioned above (A). Hep-2 cells were treated with fumonisin B1 (20 µM) or C6-ceramide (10 µg/ml), followed by deguelin (100 µM) exposure, MTT assay was used to test cell viability after 72 hours (B), Hoechst staining (C) and enzyme-linked immunosorbent cell apoptosis assay (D) were utilized to test Hep-2 cell apoptosis after 48 hours. The mean of at least three independent experiments performed in triplicate is shown. Statistical significance was analyzed by ANOVA.

    Techniques Used: MTT Assay, Staining, Apoptosis Assay

    5) Product Images from "Mutations in the SPTLC2 Subunit of Serine Palmitoyltransferase Cause Hereditary Sensory and Autonomic Neuropathy Type I"

    Article Title: Mutations in the SPTLC2 Subunit of Serine Palmitoyltransferase Cause Hereditary Sensory and Autonomic Neuropathy Type I

    Journal: American Journal of Human Genetics

    doi: 10.1016/j.ajhg.2010.09.010

    In Vitro SPT Activity Measurements of HSAN-I Associated SPTLC2 Mutants (A) Fumonisin B1 block assay. SPT activity in HEK293 cells stably expressing WT or mutant SPTLC2 is analyzed by measuring SA accumulation after treatment with Fumonisin B1. Stable expression of WT SPTLC2 generates an 8.5-fold increase in SPT activity (p = 3.24 × 10 −5 ), whereas the G382V mutant does not increase SPT activity (p = 0.18). The V359M and I504F mutations increase the activity significantly (p = 0.00063 and 0.00064, respectively) but not to the same extent as WT SPTLC2. Enhanced GFP (EGFP)-transfected cells served as control. (B) Radioactivity-based SPT activity assay. SPT activity of HEK293 cells stably expressing WT or mutant SPTLC2 was determined by measuring the incorporation of 14 ). CPM, counts per minute; SA, sphinganine. ∗∗∗ p
    Figure Legend Snippet: In Vitro SPT Activity Measurements of HSAN-I Associated SPTLC2 Mutants (A) Fumonisin B1 block assay. SPT activity in HEK293 cells stably expressing WT or mutant SPTLC2 is analyzed by measuring SA accumulation after treatment with Fumonisin B1. Stable expression of WT SPTLC2 generates an 8.5-fold increase in SPT activity (p = 3.24 × 10 −5 ), whereas the G382V mutant does not increase SPT activity (p = 0.18). The V359M and I504F mutations increase the activity significantly (p = 0.00063 and 0.00064, respectively) but not to the same extent as WT SPTLC2. Enhanced GFP (EGFP)-transfected cells served as control. (B) Radioactivity-based SPT activity assay. SPT activity of HEK293 cells stably expressing WT or mutant SPTLC2 was determined by measuring the incorporation of 14 ). CPM, counts per minute; SA, sphinganine. ∗∗∗ p

    Techniques Used: In Vitro, Single-particle Tracking, Activity Assay, Blocking Assay, Stable Transfection, Expressing, Mutagenesis, Transfection, Radioactivity

    6) Product Images from "Palmitate induced secretion of IL-6 and MCP-1 in orbital fibroblasts derived from patients with thyroid-associated ophthalmopathy"

    Article Title: Palmitate induced secretion of IL-6 and MCP-1 in orbital fibroblasts derived from patients with thyroid-associated ophthalmopathy

    Journal: Molecular Vision

    doi:

    Effect of fumonisin B1 on the palmitate-induced IL-6 and MCP-1 secretion from orbital fibroblasts. Orbital fibroblasts were pretreated with fuminosin B1 (10 μM) for 1 h, followed by treatment with palmitate (200 μM) for 24 h. Concentrations of IL-6 ( A ) and MCP-1 ( B ) were determined by ELISA. Data are presented as the mean±SD of three replicate culture wells from a representative experiment. Similar results were observed in three independent experiments using orbital fibroblasts from #1 patient with TAO ( Table 1 ). * p
    Figure Legend Snippet: Effect of fumonisin B1 on the palmitate-induced IL-6 and MCP-1 secretion from orbital fibroblasts. Orbital fibroblasts were pretreated with fuminosin B1 (10 μM) for 1 h, followed by treatment with palmitate (200 μM) for 24 h. Concentrations of IL-6 ( A ) and MCP-1 ( B ) were determined by ELISA. Data are presented as the mean±SD of three replicate culture wells from a representative experiment. Similar results were observed in three independent experiments using orbital fibroblasts from #1 patient with TAO ( Table 1 ). * p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    7) Product Images from "Short-term Mg deficiency upregulates protein kinase C isoforms in cardiovascular tissues and cells; relation to NF-kB, cytokines, ceramide salvage sphingolipid pathway and PKC-zeta: hypothesis and review"

    Article Title: Short-term Mg deficiency upregulates protein kinase C isoforms in cardiovascular tissues and cells; relation to NF-kB, cytokines, ceramide salvage sphingolipid pathway and PKC-zeta: hypothesis and review

    Journal: International Journal of Clinical and Experimental Medicine

    doi:

    [ 3 H]-palmitate and [ 3 H]-serine incorporation into free sphingosine free base in primary cultured single aortic VSM cells exposed to 0.3 mM [Mg] (with and without fumonisin B1 (FB1) - see Methods) over time; n=6-8.
    Figure Legend Snippet: [ 3 H]-palmitate and [ 3 H]-serine incorporation into free sphingosine free base in primary cultured single aortic VSM cells exposed to 0.3 mM [Mg] (with and without fumonisin B1 (FB1) - see Methods) over time; n=6-8.

    Techniques Used: Cell Culture

    8) Product Images from "Lysophosphatidylcholine as an effector of fatty acid-induced insulin resistance [S]"

    Article Title: Lysophosphatidylcholine as an effector of fatty acid-induced insulin resistance [S]

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.M014787

    Effect of ceramide synthesis inhibitors on PA-induced insulin resistance. A: L6 myotubes were pretreated with 20 μM Fumonisin B1 or 500 nM Myriocin for 1 h, then incubated with 800 μM PA for 14 h. Subsequently, L6 myotubes were treated
    Figure Legend Snippet: Effect of ceramide synthesis inhibitors on PA-induced insulin resistance. A: L6 myotubes were pretreated with 20 μM Fumonisin B1 or 500 nM Myriocin for 1 h, then incubated with 800 μM PA for 14 h. Subsequently, L6 myotubes were treated

    Techniques Used: Incubation

    9) Product Images from "Electrochemical Aptamer-Based Sensors for Rapid Point-of-Use Monitoring of the Mycotoxin Ochratoxin A Directly in a Food Stream"

    Article Title: Electrochemical Aptamer-Based Sensors for Rapid Point-of-Use Monitoring of the Mycotoxin Ochratoxin A Directly in a Food Stream

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23040912

    The E-AB ochratoxin sensor is selective enough to work when challenged directly in a realistically complex sample matrix, and highly specific for its target. ( A ) Shown here, for example, are data collected in flowing, undiluted coffee. Error bars shown are standard error of the mean for six replicate electrodes. ( B ) The E-sensor is also highly specific for its target, ochratoxin A (O. A); it does not respond to the mycotoxins, Aflatoxin B1 (A. B1), Aflatoxin B2 (A. B2), Aflatoxin G1 (A. G1), Aflatoxin G2 (A. G2), or Fumonisin B1 (F. B1). Each bar represents the KDM signal gain of sensors in buffer challenged with 10 µM of mycotoxin, error bars reflect the standard error of the mean for three replicate electrodes.
    Figure Legend Snippet: The E-AB ochratoxin sensor is selective enough to work when challenged directly in a realistically complex sample matrix, and highly specific for its target. ( A ) Shown here, for example, are data collected in flowing, undiluted coffee. Error bars shown are standard error of the mean for six replicate electrodes. ( B ) The E-sensor is also highly specific for its target, ochratoxin A (O. A); it does not respond to the mycotoxins, Aflatoxin B1 (A. B1), Aflatoxin B2 (A. B2), Aflatoxin G1 (A. G1), Aflatoxin G2 (A. G2), or Fumonisin B1 (F. B1). Each bar represents the KDM signal gain of sensors in buffer challenged with 10 µM of mycotoxin, error bars reflect the standard error of the mean for three replicate electrodes.

    Techniques Used:

    10) Product Images from "Inhibition of human tumour prostate PC-3 cell growth by cannabinoids R(+)-Methanandamide and JWH-015: Involvement of CB2"

    Article Title: Inhibition of human tumour prostate PC-3 cell growth by cannabinoids R(+)-Methanandamide and JWH-015: Involvement of CB2

    Journal: British Journal of Cancer

    doi: 10.1038/sj.bjc.6605248

    Involvement of ceramide synthesis in JWH-015-induced cell growth inhibition. ( A ) PC-3 cells were incubated in the presence of increasing concentrations of JWH-015 for 48 h and intracellular ceramide was measured by the DAG kinase method as indicated in the Methods section. ( B ) PC-3 cells were incubated with 10 μ JWH-015in the presence or absence of 2 μ SR2, 50 μ Fumonisin B1 (Fumo) or 5 μ D609 for 48 h and intracellular ceramide was assayed as above. ( C ) Cell cycle of PC-3 cells incubated with 10 μ JWH-015±50 μ Fumo for 48 h. Data are the mean±s.e. of three different experiments performed in duplicate. * P
    Figure Legend Snippet: Involvement of ceramide synthesis in JWH-015-induced cell growth inhibition. ( A ) PC-3 cells were incubated in the presence of increasing concentrations of JWH-015 for 48 h and intracellular ceramide was measured by the DAG kinase method as indicated in the Methods section. ( B ) PC-3 cells were incubated with 10 μ JWH-015in the presence or absence of 2 μ SR2, 50 μ Fumonisin B1 (Fumo) or 5 μ D609 for 48 h and intracellular ceramide was assayed as above. ( C ) Cell cycle of PC-3 cells incubated with 10 μ JWH-015±50 μ Fumo for 48 h. Data are the mean±s.e. of three different experiments performed in duplicate. * P

    Techniques Used: Inhibition, Incubation

    11) Product Images from "Electrochemical Aptamer-Based Sensors for Rapid Point-of-Use Monitoring of the Mycotoxin Ochratoxin A Directly in a Food Stream"

    Article Title: Electrochemical Aptamer-Based Sensors for Rapid Point-of-Use Monitoring of the Mycotoxin Ochratoxin A Directly in a Food Stream

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23040912

    The E-AB ochratoxin sensor is selective enough to work when challenged directly in a realistically complex sample matrix, and highly specific for its target. ( A ) Shown here, for example, are data collected in flowing, undiluted coffee. Error bars shown are standard error of the mean for six replicate electrodes. ( B ) The E-sensor is also highly specific for its target, ochratoxin A (O. A); it does not respond to the mycotoxins, Aflatoxin B1 (A. B1), Aflatoxin B2 (A. B2), Aflatoxin G1 (A. G1), Aflatoxin G2 (A. G2), or Fumonisin B1 (F. B1). Each bar represents the KDM signal gain of sensors in buffer challenged with 10 µM of mycotoxin, error bars reflect the standard error of the mean for three replicate electrodes.
    Figure Legend Snippet: The E-AB ochratoxin sensor is selective enough to work when challenged directly in a realistically complex sample matrix, and highly specific for its target. ( A ) Shown here, for example, are data collected in flowing, undiluted coffee. Error bars shown are standard error of the mean for six replicate electrodes. ( B ) The E-sensor is also highly specific for its target, ochratoxin A (O. A); it does not respond to the mycotoxins, Aflatoxin B1 (A. B1), Aflatoxin B2 (A. B2), Aflatoxin G1 (A. G1), Aflatoxin G2 (A. G2), or Fumonisin B1 (F. B1). Each bar represents the KDM signal gain of sensors in buffer challenged with 10 µM of mycotoxin, error bars reflect the standard error of the mean for three replicate electrodes.

    Techniques Used:

    12) Product Images from "Different mechanisms are involved in apoptosis induced by melanoma gangliosides on human monocyte-derived dendritic cells"

    Article Title: Different mechanisms are involved in apoptosis induced by melanoma gangliosides on human monocyte-derived dendritic cells

    Journal: Glycobiology

    doi: 10.1093/glycob/cwp015

    Ceramides generated via the de novo pathway are not responsible for DC apoptosis. ( A ) The de novo ceramides biosynthesis was blocked with Fumonisin B1 (50 μg/mL). These inhibitors were added at the beginning of cell culture in the presence or absence of gangliosides. ( B ) GM3 ganglioside induced an accumulation of ceramides via the acid SMase pathway. Acid sphingomyelinase inhibited by desipramine (1 μM) was added in the presence or absence of gangliosides every 2 days during cell culture. Results are representative of four independent experiments.
    Figure Legend Snippet: Ceramides generated via the de novo pathway are not responsible for DC apoptosis. ( A ) The de novo ceramides biosynthesis was blocked with Fumonisin B1 (50 μg/mL). These inhibitors were added at the beginning of cell culture in the presence or absence of gangliosides. ( B ) GM3 ganglioside induced an accumulation of ceramides via the acid SMase pathway. Acid sphingomyelinase inhibited by desipramine (1 μM) was added in the presence or absence of gangliosides every 2 days during cell culture. Results are representative of four independent experiments.

    Techniques Used: Generated, Cell Culture

    13) Product Images from "Plasma-Based Degradation of Mycotoxins Produced by Fusarium, Aspergillus and Alternaria Species"

    Article Title: Plasma-Based Degradation of Mycotoxins Produced by Fusarium, Aspergillus and Alternaria Species

    Journal: Toxins

    doi: 10.3390/toxins9030097

    ( a ). Time-dependent decay of four pure mycotoxins exposed to air plasma ( N = 5). (A) AAl-Toxin TA, (B) Enniatin A, (C) T2-toxin, (D) Deoxynivalenol; ( b ). Time-dependent decay of four pure mycotoxins exposed to air plasma ( N = 5). (E) Fumonisin B1, (F) Enniatin B, (G) Zearalenone, (H) Sterigmatocystin.
    Figure Legend Snippet: ( a ). Time-dependent decay of four pure mycotoxins exposed to air plasma ( N = 5). (A) AAl-Toxin TA, (B) Enniatin A, (C) T2-toxin, (D) Deoxynivalenol; ( b ). Time-dependent decay of four pure mycotoxins exposed to air plasma ( N = 5). (E) Fumonisin B1, (F) Enniatin B, (G) Zearalenone, (H) Sterigmatocystin.

    Techniques Used:

    Time-dependent decay of mycotoxins embedded in rice extract (green line) exposed to air plasma ( N = 5). ( A ) Fumonisin B1; ( B ) Enniatin B; ( C ) Zearalenone; ( D ) Sterigmatocystin. The red dotted line displays the decay slopes of the respective pure mycotoxin standards as shown in Figure 2 b.
    Figure Legend Snippet: Time-dependent decay of mycotoxins embedded in rice extract (green line) exposed to air plasma ( N = 5). ( A ) Fumonisin B1; ( B ) Enniatin B; ( C ) Zearalenone; ( D ) Sterigmatocystin. The red dotted line displays the decay slopes of the respective pure mycotoxin standards as shown in Figure 2 b.

    Techniques Used:

    14) Product Images from "Ultraviolet (UV) and Hydrogen Peroxide Activate Ceramide-ER Stress-AMPK Signaling Axis to Promote Retinal Pigment Epithelium (RPE) Cell Apoptosis"

    Article Title: Ultraviolet (UV) and Hydrogen Peroxide Activate Ceramide-ER Stress-AMPK Signaling Axis to Promote Retinal Pigment Epithelium (RPE) Cell Apoptosis

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms140510355

    UV induces ceramide production, ER stress/AMPK activation and RPE cell death. APRE-19 cells were irradiated with indicated dosage of UV; afterwards, cells were further incubated in culture medium for one hour. Phospho- and non-phospho-eIF2α/AMPKα, as well as β-actin were tested ( A ); APRE-19 cells were irradiated with UV (25 mJ/cm 2 ); afterwards, cell were incubated in culture medium for an additional 15, 30 and 60 min; cellular ceramide level was examined ( B , left panel ); APRE-19 cells were pretreated with fumonisin B1 (F-B1, 10 μM) ( B , right panel ), Sal (10 μM) or compound C (10 μM) for 1 h, followed by UV (25 mJ/cm 2 ) radiation. Cells were then cultured in culture medium for an additional 24 h; cell viability was examined by MTT assay ( C ). Experiments were repeated three times. ** p
    Figure Legend Snippet: UV induces ceramide production, ER stress/AMPK activation and RPE cell death. APRE-19 cells were irradiated with indicated dosage of UV; afterwards, cells were further incubated in culture medium for one hour. Phospho- and non-phospho-eIF2α/AMPKα, as well as β-actin were tested ( A ); APRE-19 cells were irradiated with UV (25 mJ/cm 2 ); afterwards, cell were incubated in culture medium for an additional 15, 30 and 60 min; cellular ceramide level was examined ( B , left panel ); APRE-19 cells were pretreated with fumonisin B1 (F-B1, 10 μM) ( B , right panel ), Sal (10 μM) or compound C (10 μM) for 1 h, followed by UV (25 mJ/cm 2 ) radiation. Cells were then cultured in culture medium for an additional 24 h; cell viability was examined by MTT assay ( C ). Experiments were repeated three times. ** p

    Techniques Used: Activation Assay, Irradiation, Incubation, Cell Culture, MTT Assay

    H 2 O 2 induces an early ceramide production, inhibited by fumonisin B1. APRE-19 cells ( A , C ) or primary mouse RPE cells ( D , E ) were either left untreated or treated with H 2 O 2 (200 μM) or 5-amino-1-β-D-ribofuranosyl-imidazole-4-carboxamide (AICAR) (1 mM) or indicated time points; cellular ceramide production was analyzed, quantified and was expressed as fold changes vs. untreated control group; phospho- and non-phospho- AMPK, ACC and eIF2α were detected as described above ( E ); ( B ) APRE-19 cells were pretreated with ceramide de novo synthase inhibitor fumonisin B1 (10 μM) for one hour, followed by H 2 O 2 (200 μM) for indicated time points; cellular ceramides production was analyzed, quantified and expressed as fold changes vs. control group (Ctrl). Experiments were repeated three times. * p
    Figure Legend Snippet: H 2 O 2 induces an early ceramide production, inhibited by fumonisin B1. APRE-19 cells ( A , C ) or primary mouse RPE cells ( D , E ) were either left untreated or treated with H 2 O 2 (200 μM) or 5-amino-1-β-D-ribofuranosyl-imidazole-4-carboxamide (AICAR) (1 mM) or indicated time points; cellular ceramide production was analyzed, quantified and was expressed as fold changes vs. untreated control group; phospho- and non-phospho- AMPK, ACC and eIF2α were detected as described above ( E ); ( B ) APRE-19 cells were pretreated with ceramide de novo synthase inhibitor fumonisin B1 (10 μM) for one hour, followed by H 2 O 2 (200 μM) for indicated time points; cellular ceramides production was analyzed, quantified and expressed as fold changes vs. control group (Ctrl). Experiments were repeated three times. * p

    Techniques Used:

    The proposed signaling pathway of this study. In RPE cells, UV radiation induces ROS production to induce an early ceramide production. Increased ceramide activates ER stress, which serves as an upstream signaling for AMPK activation. AMPK activation appears to be pro-apoptotic in this system. Suppression of this signaling axis by ceramide synthase inhibitor fumonisin B1, ER stress inhibitor salubrinal or by AMPK inhibitor compound C inhibits UV or H 2 O 2 -induced RPE cell death, while C6 ceramide and AMPK activator AICAR mimicked UV/H 2 O 2 ’s effect. The role of MAPK activation in UV or H 2 O 2 -induced RPE cell death needs further investigation; also, the mechanism link between these pathways warrants more studies.
    Figure Legend Snippet: The proposed signaling pathway of this study. In RPE cells, UV radiation induces ROS production to induce an early ceramide production. Increased ceramide activates ER stress, which serves as an upstream signaling for AMPK activation. AMPK activation appears to be pro-apoptotic in this system. Suppression of this signaling axis by ceramide synthase inhibitor fumonisin B1, ER stress inhibitor salubrinal or by AMPK inhibitor compound C inhibits UV or H 2 O 2 -induced RPE cell death, while C6 ceramide and AMPK activator AICAR mimicked UV/H 2 O 2 ’s effect. The role of MAPK activation in UV or H 2 O 2 -induced RPE cell death needs further investigation; also, the mechanism link between these pathways warrants more studies.

    Techniques Used: Activation Assay

    H 2 O 2 -induced RPE cell apoptosis is suppressed by ceramide-ER stress-AMPK inhibitors. APRE-19 cells were pretreated with Sal (10 μM) for one hour, followed by H 2 O 2 (200 μM) for 24 h; cell apoptosis was detected by TUNEL staining ( A , B ); Cultured APRE-19 cells (RPE cells) were treated as follows: control (Ctrl), H 2 O 2 (200 μM), fumonisin B1 (F-B1, 10 μM), compound C (10 μM), fumonisin B1 + H 2 O 2 , compound C + H 2 O 2 ( C , D ), AICAR (1 mM) and C6-ceramide (10 μg/mL) ( E ); cell apoptosis was detected by TUNEL staining. Experiments were repeated three times. * p
    Figure Legend Snippet: H 2 O 2 -induced RPE cell apoptosis is suppressed by ceramide-ER stress-AMPK inhibitors. APRE-19 cells were pretreated with Sal (10 μM) for one hour, followed by H 2 O 2 (200 μM) for 24 h; cell apoptosis was detected by TUNEL staining ( A , B ); Cultured APRE-19 cells (RPE cells) were treated as follows: control (Ctrl), H 2 O 2 (200 μM), fumonisin B1 (F-B1, 10 μM), compound C (10 μM), fumonisin B1 + H 2 O 2 , compound C + H 2 O 2 ( C , D ), AICAR (1 mM) and C6-ceramide (10 μg/mL) ( E ); cell apoptosis was detected by TUNEL staining. Experiments were repeated three times. * p

    Techniques Used: TUNEL Assay, Staining, Cell Culture

    15) Product Images from "Cellular and Molecular Mechanisms Mediated by recPrPC Involved in the Neuronal Differentiation Process of Mesenchymal Stem Cells"

    Article Title: Cellular and Molecular Mechanisms Mediated by recPrPC Involved in the Neuronal Differentiation Process of Mesenchymal Stem Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20020345

    Effect of lipid rafts perturbation on Akt and ERK Phosphorylation induced by recPrP C . hDPSCs, untreated or treated with 0.5 µg/mL of recPrP C for 10 min in the presence or in the absence of Fumonisin B1 or MβCD, were analyzed by Western blot using anti-pAkt and anti-total Akt ( A ). anti-pERK1/2 and anti-total ERK1/2 ( B ). Densitometric analysis is shown in the right panel. Results represent the mean ± SD from 3 independent experiments, * p recPrP C treated cells
    Figure Legend Snippet: Effect of lipid rafts perturbation on Akt and ERK Phosphorylation induced by recPrP C . hDPSCs, untreated or treated with 0.5 µg/mL of recPrP C for 10 min in the presence or in the absence of Fumonisin B1 or MβCD, were analyzed by Western blot using anti-pAkt and anti-total Akt ( A ). anti-pERK1/2 and anti-total ERK1/2 ( B ). Densitometric analysis is shown in the right panel. Results represent the mean ± SD from 3 independent experiments, * p recPrP C treated cells

    Techniques Used: Western Blot

    16) Product Images from "Staurosporines decrease ORMDL proteins and enhance sphingomyelin synthesis resulting in depletion of plasmalemmal phosphatidylserine"

    Article Title: Staurosporines decrease ORMDL proteins and enhance sphingomyelin synthesis resulting in depletion of plasmalemmal phosphatidylserine

    Journal: Scientific Reports

    doi: 10.1038/srep35762

    Staurosporine increases sphingomyelin contents in both the exofacial leaflet of the plasma membrane and intracellular compartments. ( a ) Confocal images of CHO cells treated with DMSO, 50 nM STS or 50 nM STS + 15 μM fumonisin B1 (FB1) for 24 hr labelled with MBP-GFP-NT-Lysenin (Lys) protein. Bar, 10 μm. ( b ) Binding of MBP-GFP-NT-Lys protein to the exofacial leaflets of the plasma membrane in DMSO, STS or STS + FB1 treated CHO cells. Data are means ± SEM (n = 3). ***p
    Figure Legend Snippet: Staurosporine increases sphingomyelin contents in both the exofacial leaflet of the plasma membrane and intracellular compartments. ( a ) Confocal images of CHO cells treated with DMSO, 50 nM STS or 50 nM STS + 15 μM fumonisin B1 (FB1) for 24 hr labelled with MBP-GFP-NT-Lysenin (Lys) protein. Bar, 10 μm. ( b ) Binding of MBP-GFP-NT-Lys protein to the exofacial leaflets of the plasma membrane in DMSO, STS or STS + FB1 treated CHO cells. Data are means ± SEM (n = 3). ***p

    Techniques Used: Binding Assay

    Determination of the molecular species of sphingomyelin following low-dose staurosporine. Quantitative lipidomic analysis of sphingomyelin species from control CHO cells and treated with 50 nM staurosporine (STS), 50 nM 7-oxostaurosporine (OSS), 15 μM fumonisin B1 (FB1) and 50 nM STS + 15 μM FB1 for 24 hours. Lipid composition is shown normalized to protein to compare the changes due to incubation with the drugs ( a ) and then data are normalized by the value of DMSO samples ( b ). Data are means ± SEM (n = 4). *p
    Figure Legend Snippet: Determination of the molecular species of sphingomyelin following low-dose staurosporine. Quantitative lipidomic analysis of sphingomyelin species from control CHO cells and treated with 50 nM staurosporine (STS), 50 nM 7-oxostaurosporine (OSS), 15 μM fumonisin B1 (FB1) and 50 nM STS + 15 μM FB1 for 24 hours. Lipid composition is shown normalized to protein to compare the changes due to incubation with the drugs ( a ) and then data are normalized by the value of DMSO samples ( b ). Data are means ± SEM (n = 4). *p

    Techniques Used: Incubation

    Inhibition of SM synthesis by fumonisin B1 prevents the relocalization of PtdSer. ( a ) Confocal images of CHO cells treated with DMSO, 50 nM STS or 50 nM STS + 15 μM FB1 for 24 hr expressing GFP-Lact-C2 and mCherry-D4H. Bar, 10 μm. ( b ) Quantitation of fluorescence signal plasma membrane:cytoplasm of GFP-Lact-C2 and mCherry-D4H in ( a ). Total 50 cells from three independent experiments were analysed. Data are means ± SEM. **p
    Figure Legend Snippet: Inhibition of SM synthesis by fumonisin B1 prevents the relocalization of PtdSer. ( a ) Confocal images of CHO cells treated with DMSO, 50 nM STS or 50 nM STS + 15 μM FB1 for 24 hr expressing GFP-Lact-C2 and mCherry-D4H. Bar, 10 μm. ( b ) Quantitation of fluorescence signal plasma membrane:cytoplasm of GFP-Lact-C2 and mCherry-D4H in ( a ). Total 50 cells from three independent experiments were analysed. Data are means ± SEM. **p

    Techniques Used: Inhibition, Expressing, Quantitation Assay, Fluorescence

    17) Product Images from "Intrinsic, Pro-Apoptotic Effects of IGFBP-3 on Breast Cancer Cells are Reversible: Involvement of PKA, Rho, and Ceramide"

    Article Title: Intrinsic, Pro-Apoptotic Effects of IGFBP-3 on Breast Cancer Cells are Reversible: Involvement of PKA, Rho, and Ceramide

    Journal: Frontiers in Endocrinology

    doi: 10.3389/fendo.2011.00013

    Hs578T were seeded at 0.3 × 10 6 per T25 culture flasks for 24 h after which the GM was replaced with SFM for 24 h. Cells were treated with IGFBP-3 (100 ng/ml) for 30 min following a 30-min pre-dose with fumonisin B1 (1 μM) . (A) Shows Western immunoblots of p-MAPK and ERK-2 and are representative of experiments repeated three times. (B) Shows a graph of the mean arbitrary optical density measurements from three experiments demonstrating changes in p-MAPK corrected for total ERK-2. (C) Hs578T cells were seeded in six-well plates at 0.1 × 10 6 cells in GM followed by 24 h in SFM prior to pre-dosing for 24 h with IGFBP-5 (100 ng/ml) ± fumonisin B1 (1 μM). Cells were then re-dosed for 24 h as above ± an apoptotic dose of C2-ceramide. The graph represents changes in cell death and shows the mean ± SEM of three experiments each repeated in triplicate.
    Figure Legend Snippet: Hs578T were seeded at 0.3 × 10 6 per T25 culture flasks for 24 h after which the GM was replaced with SFM for 24 h. Cells were treated with IGFBP-3 (100 ng/ml) for 30 min following a 30-min pre-dose with fumonisin B1 (1 μM) . (A) Shows Western immunoblots of p-MAPK and ERK-2 and are representative of experiments repeated three times. (B) Shows a graph of the mean arbitrary optical density measurements from three experiments demonstrating changes in p-MAPK corrected for total ERK-2. (C) Hs578T cells were seeded in six-well plates at 0.1 × 10 6 cells in GM followed by 24 h in SFM prior to pre-dosing for 24 h with IGFBP-5 (100 ng/ml) ± fumonisin B1 (1 μM). Cells were then re-dosed for 24 h as above ± an apoptotic dose of C2-ceramide. The graph represents changes in cell death and shows the mean ± SEM of three experiments each repeated in triplicate.

    Techniques Used: Western Blot

    Hs578T cells were seeded in six-well plates at 0.1 × 10 6 cells in GM followed by 24 h in SFM prior to pre-dosing for 24 h (A) with IGFBP-3 (100 ng/ml), (B) SPD (10 ng/ml) or (C) IGFBP-3 ± fumonisin B1 (1 μM) . Cells were then re-dosed for 24 h ± an apoptotic dose of C2-ceramide (A,B) or antimycin A (C) . The graphs represent changes in cell death and show the mean ± SEM of three experiments each repeated in triplicate.
    Figure Legend Snippet: Hs578T cells were seeded in six-well plates at 0.1 × 10 6 cells in GM followed by 24 h in SFM prior to pre-dosing for 24 h (A) with IGFBP-3 (100 ng/ml), (B) SPD (10 ng/ml) or (C) IGFBP-3 ± fumonisin B1 (1 μM) . Cells were then re-dosed for 24 h ± an apoptotic dose of C2-ceramide (A,B) or antimycin A (C) . The graphs represent changes in cell death and show the mean ± SEM of three experiments each repeated in triplicate.

    Techniques Used:

    18) Product Images from "Sphingosine Kinase Isoforms Regulate Oxaliplatin Sensitivity of Human Colon Cancer Cells through Ceramide Accumulation and Akt Activation"

    Article Title: Sphingosine Kinase Isoforms Regulate Oxaliplatin Sensitivity of Human Colon Cancer Cells through Ceramide Accumulation and Akt Activation

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M900735200

    Effects of various inhibitors on l -OHP-induced ceramide formation, caspase activity, PARP degradation, and cell viability in HCT116 cells. HCT116 cells were pretreated with or without fumonisin B1 ( FB1 ; 100 μg/ml), neutral SMase inhibitor GW4869
    Figure Legend Snippet: Effects of various inhibitors on l -OHP-induced ceramide formation, caspase activity, PARP degradation, and cell viability in HCT116 cells. HCT116 cells were pretreated with or without fumonisin B1 ( FB1 ; 100 μg/ml), neutral SMase inhibitor GW4869

    Techniques Used: Activity Assay

    19) Product Images from "Resveratrol suppresses growth of cancer stem-like cells by inhibiting fatty acid synthase"

    Article Title: Resveratrol suppresses growth of cancer stem-like cells by inhibiting fatty acid synthase

    Journal: Breast Cancer Research and Treatment

    doi: 10.1007/s10549-010-1300-6

    Resveratrol activates pro-apoptotic genes in tumor stem-like cells A Tumor stem-like cell were isolated from MDA-MB231 cells and treated with or without resveratrol in the presence or absence of TOFA or Fumonisin B1 for 48 hours followed by assaying for the apoptotic index by In situ cell death detection kit/TMR red. B, C, D Tumor stem-like cells were isolated from MCF7 and MDA-MB231 cells and incubated with various concentrations of resveratrol for 48 hours followed by quantifying the mRNA expression of the pro-apoptotic genes DAPK2, BNIP3 and p21 by qRT-PCR.
    Figure Legend Snippet: Resveratrol activates pro-apoptotic genes in tumor stem-like cells A Tumor stem-like cell were isolated from MDA-MB231 cells and treated with or without resveratrol in the presence or absence of TOFA or Fumonisin B1 for 48 hours followed by assaying for the apoptotic index by In situ cell death detection kit/TMR red. B, C, D Tumor stem-like cells were isolated from MCF7 and MDA-MB231 cells and incubated with various concentrations of resveratrol for 48 hours followed by quantifying the mRNA expression of the pro-apoptotic genes DAPK2, BNIP3 and p21 by qRT-PCR.

    Techniques Used: Isolation, Multiple Displacement Amplification, In Situ, Incubation, Expressing, Quantitative RT-PCR

    20) Product Images from "Lipid Rafts in the Maintenance of Synapses, Dendritic Spines, and Surface AMPA Receptor Stability"

    Article Title: Lipid Rafts in the Maintenance of Synapses, Dendritic Spines, and Surface AMPA Receptor Stability

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.23-08-03262.2003

    Detergent-resistant membranes in dendrites of cultured neurons. A , Hippocampal neuron at 21 DIV double-labeled with DiIC 18 (red) and FAST-DiO (green). B , DiIC 18 /FAST-DiO-labeled hippocampal neuron after extraction with 0.5% Triton X-100 at 4°C. C , Hippocampal neuron treated with mevastatin, fumonisin B 1 , and mevalonate for 5 d labeled with DiIC 18 and FAST-DiO. D , DiIC 18 /FAST-DiO-labeled hippocampal neuron treated as in C, after extraction with 0.5% Triton X-100 at 4°C. Scale bar, 20 μm. E , Cholesterol and sphingolipid levels in control and raft-depleted cultures. Cholesterol levels in raft-depleted cultures were decreased ∼31% ( p
    Figure Legend Snippet: Detergent-resistant membranes in dendrites of cultured neurons. A , Hippocampal neuron at 21 DIV double-labeled with DiIC 18 (red) and FAST-DiO (green). B , DiIC 18 /FAST-DiO-labeled hippocampal neuron after extraction with 0.5% Triton X-100 at 4°C. C , Hippocampal neuron treated with mevastatin, fumonisin B 1 , and mevalonate for 5 d labeled with DiIC 18 and FAST-DiO. D , DiIC 18 /FAST-DiO-labeled hippocampal neuron treated as in C, after extraction with 0.5% Triton X-100 at 4°C. Scale bar, 20 μm. E , Cholesterol and sphingolipid levels in control and raft-depleted cultures. Cholesterol levels in raft-depleted cultures were decreased ∼31% ( p

    Techniques Used: Cell Culture, Labeling

    Lipid-raft depletion reduces the number but increases the size of synapses. All images were taken from hippocampal neurons at 21 DIV. A1 , Control hippocampal neurons stained for PSD-95. A2 , Hippocampal neurons treated for 7 d with mevastatin, fumonisin B 1 , and mevalonate (raft-depleted) and stained for PSD-95. A3 , Quantitation of PSD-95 cluster size and density in control (ctrl) and raft-depleted (depl) neurons. Histograms show mean ± SEM; n = 5 microscope fields for each condition. Differences in cluster size ( p = 0.0036) and density ( p = 0.0024) are significant (Student's t test). B–E , Control and raft-depleted neurons stained for NR1 subunit of NMDA receptor (NMDAR; B1 , B2 ), Shank ( C1 , C2 ), bassoon (D1, D2 ), and β2/3 subunit of GABA A receptor (GABA A R; E1 , E2 ). Scale bar, 20 μm. F , Total cell lysate of control and raft-depleted hippocampal cultures immunoblotted for the indicated proteins. Anti-tubulin immunoblotting confirmed equal loading of protein.
    Figure Legend Snippet: Lipid-raft depletion reduces the number but increases the size of synapses. All images were taken from hippocampal neurons at 21 DIV. A1 , Control hippocampal neurons stained for PSD-95. A2 , Hippocampal neurons treated for 7 d with mevastatin, fumonisin B 1 , and mevalonate (raft-depleted) and stained for PSD-95. A3 , Quantitation of PSD-95 cluster size and density in control (ctrl) and raft-depleted (depl) neurons. Histograms show mean ± SEM; n = 5 microscope fields for each condition. Differences in cluster size ( p = 0.0036) and density ( p = 0.0024) are significant (Student's t test). B–E , Control and raft-depleted neurons stained for NR1 subunit of NMDA receptor (NMDAR; B1 , B2 ), Shank ( C1 , C2 ), bassoon (D1, D2 ), and β2/3 subunit of GABA A receptor (GABA A R; E1 , E2 ). Scale bar, 20 μm. F , Total cell lysate of control and raft-depleted hippocampal cultures immunoblotted for the indicated proteins. Anti-tubulin immunoblotting confirmed equal loading of protein.

    Techniques Used: Staining, Quantitation Assay, Microscopy

    21) Product Images from "Fatty acid-induced ? cell apoptosis: A link between obesity and diabetes"

    Article Title: Fatty acid-induced ? cell apoptosis: A link between obesity and diabetes

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

    doi:

    ( A ) Effect of FFA on DNA fragmentation in pancreatic islets from lean wild-type (+/+) and obese prediabetic homozygous ( fa/fa ) ZDF rats. Pancreatic islets were isolated from 7-week-old rats and cultured with 1 mM FFA as indicated. Ladder %, fragmented DNA percent. ( B ) Effect of FFA on ceramide content in pancreatic islets of ZDF rats. Islets from lean wild-type (•) and obese homozygous (□) ZDF rats were cultured with 0 or 1 mM FFA at the indicated times, and ceramide contents were determined. ( C ) De novo synthesis of [ 3 H]ceramide from [ 3 H]palmitate in islets of wild type (+/+) and fa/fa ZDF rats in the absence and presence of the ceramide synthase inhibitor fumonisin B 1 (FB 1 ). Rats were 7 weeks of age. Values are the mean ± SEM of triplicate experiments. ( D ) Comparison of [ 3 H]ceramide and [ 3 H]H 2 O to assess relative rates of de novo ceramide synthesis vs. oxidation of [ 3 H]palmitate by isolated islets of +/+ and fa/fa rats. Values are the mean ± SEM of triplicate experiments. ∗, P
    Figure Legend Snippet: ( A ) Effect of FFA on DNA fragmentation in pancreatic islets from lean wild-type (+/+) and obese prediabetic homozygous ( fa/fa ) ZDF rats. Pancreatic islets were isolated from 7-week-old rats and cultured with 1 mM FFA as indicated. Ladder %, fragmented DNA percent. ( B ) Effect of FFA on ceramide content in pancreatic islets of ZDF rats. Islets from lean wild-type (•) and obese homozygous (□) ZDF rats were cultured with 0 or 1 mM FFA at the indicated times, and ceramide contents were determined. ( C ) De novo synthesis of [ 3 H]ceramide from [ 3 H]palmitate in islets of wild type (+/+) and fa/fa ZDF rats in the absence and presence of the ceramide synthase inhibitor fumonisin B 1 (FB 1 ). Rats were 7 weeks of age. Values are the mean ± SEM of triplicate experiments. ( D ) Comparison of [ 3 H]ceramide and [ 3 H]H 2 O to assess relative rates of de novo ceramide synthesis vs. oxidation of [ 3 H]palmitate by isolated islets of +/+ and fa/fa rats. Values are the mean ± SEM of triplicate experiments. ∗, P

    Techniques Used: Isolation, Cell Culture

    Effect of exogenous ceramide and of blockade of ceramide synthesis on DNA fragmentation in islets from obese fa/fa ZDF rats. Islets isolated from 7-week-old obese prediabetic fa/fa ZDF rats were cultured for 24 hr in medium containing 15 μM C 2 -ceramide (C2-Cer) without FFA or 1 mM FFA plus 50 μM fumonisin B 1 (FB 1 ), an inhibitor of ceramide synthetase. Lane 1, 100-bp DNA size marker (Boeringer Mannheim); Ladder (%), fragmented DNA percent.
    Figure Legend Snippet: Effect of exogenous ceramide and of blockade of ceramide synthesis on DNA fragmentation in islets from obese fa/fa ZDF rats. Islets isolated from 7-week-old obese prediabetic fa/fa ZDF rats were cultured for 24 hr in medium containing 15 μM C 2 -ceramide (C2-Cer) without FFA or 1 mM FFA plus 50 μM fumonisin B 1 (FB 1 ), an inhibitor of ceramide synthetase. Lane 1, 100-bp DNA size marker (Boeringer Mannheim); Ladder (%), fragmented DNA percent.

    Techniques Used: Isolation, Cell Culture, Marker

    22) Product Images from "Doxorubicin blocks proliferation of cancer cells through proteolytic activation of CREB3L1Decision letterAuthor response"

    Article Title: Doxorubicin blocks proliferation of cancer cells through proteolytic activation of CREB3L1Decision letterAuthor response

    Journal: eLife

    doi: 10.7554/eLife.00090.011

    Doxorubicin-induced synthesis of ceramide stimulates cleavage of CREB3L1. ( A ),( C ) On day 0, Huh7 cells were seeded at 4 × 10 5 per 60-mm dish. On day 1, the cells were treated with indicated concentrations of myriocin ( A ) or fumonisin B1 ( C ) for 2 hr, followed by co-incubation with 200 nM doxorubicin. On day 2, 24 hr after the doxorubicin treatment, cells were analyzed for cleavage of CREB3L1 by immunoblot analysis as described in Figure 1B . ( B ) Huh7 cells treated with or without 30 µM myriocin for 2 hr followed by co-treatment with doxorubicin were analyzed as described in Figure 3A . ( D ) Huh7 cells treated with 10 µM C 6 -ceramide for 3 hr were analyzed as described in Figure 6B . Results are reported as mean ± S.E.M. of triplicate incubations from a representative experiment. ( E ) Huh7 cells treated with indicated concentration of C 6 -ceramide for 24 hr were analyzed as described in Figure 1B . ( F ) Indicated cells treated with indicated concentration of C 6 -ceramide for 48 hr were analyzed as described in Figure 3A . ( B ),( F ) Results are reported as mean ± S.E.M. of three independent experiments. DOI: http://dx.doi.org/10.7554/eLife.00090.009
    Figure Legend Snippet: Doxorubicin-induced synthesis of ceramide stimulates cleavage of CREB3L1. ( A ),( C ) On day 0, Huh7 cells were seeded at 4 × 10 5 per 60-mm dish. On day 1, the cells were treated with indicated concentrations of myriocin ( A ) or fumonisin B1 ( C ) for 2 hr, followed by co-incubation with 200 nM doxorubicin. On day 2, 24 hr after the doxorubicin treatment, cells were analyzed for cleavage of CREB3L1 by immunoblot analysis as described in Figure 1B . ( B ) Huh7 cells treated with or without 30 µM myriocin for 2 hr followed by co-treatment with doxorubicin were analyzed as described in Figure 3A . ( D ) Huh7 cells treated with 10 µM C 6 -ceramide for 3 hr were analyzed as described in Figure 6B . Results are reported as mean ± S.E.M. of triplicate incubations from a representative experiment. ( E ) Huh7 cells treated with indicated concentration of C 6 -ceramide for 24 hr were analyzed as described in Figure 1B . ( F ) Indicated cells treated with indicated concentration of C 6 -ceramide for 48 hr were analyzed as described in Figure 3A . ( B ),( F ) Results are reported as mean ± S.E.M. of three independent experiments. DOI: http://dx.doi.org/10.7554/eLife.00090.009

    Techniques Used: Incubation, Concentration Assay

    23) Product Images from "Short-term magnesium deficiency upregulates sphingomyelin synthase and p53 in cardiovascular tissues and cells: relevance to the de novo synthesis of ceramide"

    Article Title: Short-term magnesium deficiency upregulates sphingomyelin synthase and p53 in cardiovascular tissues and cells: relevance to the de novo synthesis of ceramide

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    doi: 10.1152/ajpheart.00671.2010

    Influence of fumonisin B1 and myriocin (ISP-1) on [ 3 H]palmitic acid incorporation into CVSM and AVSM as a function of [Mg 2+ ] o during either a 3- or 18-h incubation period. Values are means ± SE; n = 10–12 animals/group. *Experimental mean values are significantly different from their respective paired mean control values ( P
    Figure Legend Snippet: Influence of fumonisin B1 and myriocin (ISP-1) on [ 3 H]palmitic acid incorporation into CVSM and AVSM as a function of [Mg 2+ ] o during either a 3- or 18-h incubation period. Values are means ± SE; n = 10–12 animals/group. *Experimental mean values are significantly different from their respective paired mean control values ( P

    Techniques Used: Incubation

    24) Product Images from "Prostaglandin E2 suppresses the differentiation of retinoic acid-producing dendritic cells in mice and humans"

    Article Title: Prostaglandin E2 suppresses the differentiation of retinoic acid-producing dendritic cells in mice and humans

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20101967

    Stroma-derived PGE2 inhibits RALDH expression during DC differentiation. (a) BM-DCs were differentiated with GM-CSF in the presence of skin stroma SN that had been heated at 80°C or digested with trypsin, or SN derived from skin stroma grown in the presence of fumonisin B1 (FB1), indomethacin (indo), or NBDJ. After 3 d, LPS was added and, 18 h later, cells were stained with ALD and CD11c to analyze DC RALDH activity. Bar graphs show the mean relative inhibition of ALD + CD11c + DCs between treated SN and untreated parent SN with SD. Relative inhibition was calculated as described in Materials and methods. Values were pooled from 2–10 experiments. (b) Pathway of prostaglandin synthesis. (c) BM cells were cultured with GM-CSF alone or in the presence of SN from WT or indomethacin-treated skin stroma (SN at 25% culture volume). 0.1 µM of exogenous individual prostaglandins subtypes was added at the beginning of culture. After 3 d, LPS was added and, 18 h later, cells were stained with ALD and CD11c to analyze DC RALDH activity. A representative contour plot of CD11c versus ALD staining is shown with inset values of the mean percentage of CD11c + DCs that are ALD + pooled from three experiments. (d) The concentration of PGE2 in skin stromal SN was determined by ELISA. Shown is the mean with SD from seven individual lines.
    Figure Legend Snippet: Stroma-derived PGE2 inhibits RALDH expression during DC differentiation. (a) BM-DCs were differentiated with GM-CSF in the presence of skin stroma SN that had been heated at 80°C or digested with trypsin, or SN derived from skin stroma grown in the presence of fumonisin B1 (FB1), indomethacin (indo), or NBDJ. After 3 d, LPS was added and, 18 h later, cells were stained with ALD and CD11c to analyze DC RALDH activity. Bar graphs show the mean relative inhibition of ALD + CD11c + DCs between treated SN and untreated parent SN with SD. Relative inhibition was calculated as described in Materials and methods. Values were pooled from 2–10 experiments. (b) Pathway of prostaglandin synthesis. (c) BM cells were cultured with GM-CSF alone or in the presence of SN from WT or indomethacin-treated skin stroma (SN at 25% culture volume). 0.1 µM of exogenous individual prostaglandins subtypes was added at the beginning of culture. After 3 d, LPS was added and, 18 h later, cells were stained with ALD and CD11c to analyze DC RALDH activity. A representative contour plot of CD11c versus ALD staining is shown with inset values of the mean percentage of CD11c + DCs that are ALD + pooled from three experiments. (d) The concentration of PGE2 in skin stromal SN was determined by ELISA. Shown is the mean with SD from seven individual lines.

    Techniques Used: Derivative Assay, Expressing, Staining, Activity Assay, Inhibition, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

    25) Product Images from "Cellular and Molecular Mechanisms Mediated by recPrPC Involved in the Neuronal Differentiation Process of Mesenchymal Stem Cells"

    Article Title: Cellular and Molecular Mechanisms Mediated by recPrPC Involved in the Neuronal Differentiation Process of Mesenchymal Stem Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20020345

    Effect of lipid rafts perturbation on Akt and ERK Phosphorylation induced by recPrP C . hDPSCs, untreated or treated with 0.5 µg/mL of recPrP C for 10 min in the presence or in the absence of Fumonisin B1 or MβCD, were analyzed by Western blot using anti-pAkt and anti-total Akt ( A ). anti-pERK1/2 and anti-total ERK1/2 ( B ). Densitometric analysis is shown in the right panel. Results represent the mean ± SD from 3 independent experiments, * p recPrP C treated cells
    Figure Legend Snippet: Effect of lipid rafts perturbation on Akt and ERK Phosphorylation induced by recPrP C . hDPSCs, untreated or treated with 0.5 µg/mL of recPrP C for 10 min in the presence or in the absence of Fumonisin B1 or MβCD, were analyzed by Western blot using anti-pAkt and anti-total Akt ( A ). anti-pERK1/2 and anti-total ERK1/2 ( B ). Densitometric analysis is shown in the right panel. Results represent the mean ± SD from 3 independent experiments, * p recPrP C treated cells

    Techniques Used: Western Blot

    26) Product Images from "Peroxidase-dependent apoplastic oxidative burst in Arabidopsis required for pathogen resistance"

    Article Title: Peroxidase-dependent apoplastic oxidative burst in Arabidopsis required for pathogen resistance

    Journal: The Plant journal : for cell and molecular biology

    doi: 10.1111/j.1365-313X.2006.02837.x

    Susceptibility of FBP1 transgenic lines to the fungal toxin fumonisin B1
    Figure Legend Snippet: Susceptibility of FBP1 transgenic lines to the fungal toxin fumonisin B1

    Techniques Used: Transgenic Assay

    27) Product Images from "Curcumin alleviates diabetic nephropathy via inhibiting podocyte mesenchymal transdifferentiation and inducing autophagy in rats and MPC5 cells"

    Article Title: Curcumin alleviates diabetic nephropathy via inhibiting podocyte mesenchymal transdifferentiation and inducing autophagy in rats and MPC5 cells

    Journal: Pharmaceutical Biology

    doi: 10.1080/13880209.2019.1688843

    Effect of curcumin on E-cadherin, vimentin and TWIST1 proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of E-cadherin, vimentin and TWIST1 protein from MPC5 cells were detected by Western blot and normalized to β-actin and then (B–D) relative band intensities were used in order to quantify E-cadherin, vimentin and TWIST1 protein. * p
    Figure Legend Snippet: Effect of curcumin on E-cadherin, vimentin and TWIST1 proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of E-cadherin, vimentin and TWIST1 protein from MPC5 cells were detected by Western blot and normalized to β-actin and then (B–D) relative band intensities were used in order to quantify E-cadherin, vimentin and TWIST1 protein. * p

    Techniques Used: Cell Culture, Western Blot

    Effect of curcumin on PI3K and p-Akt proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of PI3K and p-Akt proteins from renal tissues were detected by Western blot and normalized to β-actin and then (B–C) relative band intensities were used in order to quantify PI3K and p-Akt proteins. * p
    Figure Legend Snippet: Effect of curcumin on PI3K and p-Akt proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of PI3K and p-Akt proteins from renal tissues were detected by Western blot and normalized to β-actin and then (B–C) relative band intensities were used in order to quantify PI3K and p-Akt proteins. * p

    Techniques Used: Cell Culture, Western Blot

    Effect of curcumin on p62, LC3 and p-mTOR proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of p62, LC3 and p-mTOR proteins from renal tissues were detected by Western blot and normalized to β-actin and then (B–D) relative band intensities were used in order to quantify p62, LC3 and p-mTOR proteins. * p
    Figure Legend Snippet: Effect of curcumin on p62, LC3 and p-mTOR proteins in MPC5 cells. (A) EMT-induced MPC5 cells were established and cultured in curcumin with or with fumonisin B1 or 3BDO. The levels of p62, LC3 and p-mTOR proteins from renal tissues were detected by Western blot and normalized to β-actin and then (B–D) relative band intensities were used in order to quantify p62, LC3 and p-mTOR proteins. * p

    Techniques Used: Cell Culture, Western Blot

    28) Product Images from "Phospholipid Composition of Purified Chlamydia trachomatis Mimics That of the Eucaryotic Host Cell"

    Article Title: Phospholipid Composition of Purified Chlamydia trachomatis Mimics That of the Eucaryotic Host Cell

    Journal: Infection and Immunity

    doi:

    Summary of the effects of brefeldin A, fumonisin B 1 , and sphingomyelinase (SMase) treatment on SM content of mock-infected mouse L cells (open bars), C. trachomatis -infected mouse L cells at 40 h p.i. (upward hatched bars), and highly purified EBs prepared from infected mouse L cells at 40 h p.i. (downward hatched bars).
    Figure Legend Snippet: Summary of the effects of brefeldin A, fumonisin B 1 , and sphingomyelinase (SMase) treatment on SM content of mock-infected mouse L cells (open bars), C. trachomatis -infected mouse L cells at 40 h p.i. (upward hatched bars), and highly purified EBs prepared from infected mouse L cells at 40 h p.i. (downward hatched bars).

    Techniques Used: Infection, Purification

    Incorporation of [ 14 C]isoleucine into glycerophospholipids of C. trachomatis -infected mouse L cells in the absence (open bars) and presence (hatched bars) of 20 μM fumonisin B 1 . Fumonisin B 1 was added at the start of the infection (2 h p.i.), and radiolabelled isoleucine was added at 20 h p.i. The cells were harvested 5 h later. Results are expressed as means ± standard deviations of results from three separate experiments. CL, cardiolipin.
    Figure Legend Snippet: Incorporation of [ 14 C]isoleucine into glycerophospholipids of C. trachomatis -infected mouse L cells in the absence (open bars) and presence (hatched bars) of 20 μM fumonisin B 1 . Fumonisin B 1 was added at the start of the infection (2 h p.i.), and radiolabelled isoleucine was added at 20 h p.i. The cells were harvested 5 h later. Results are expressed as means ± standard deviations of results from three separate experiments. CL, cardiolipin.

    Techniques Used: Infection

    Incorporation of [ 14 C]serine into phospholipids of mock-infected mouse L cells (A) and C. trachomatis -infected mouse L cells (B) in the absence (open bars) and presence (hatched bars) of 20 μM fumonisin B 1 . Fumonisin B 1 was added at the start of the infection (2 h p.i.), and radiolabelled serine was added at 20 h p.i. The cells were harvested 5 h later. Results are expressed as means ± standard deviations of results from three separate experiments.
    Figure Legend Snippet: Incorporation of [ 14 C]serine into phospholipids of mock-infected mouse L cells (A) and C. trachomatis -infected mouse L cells (B) in the absence (open bars) and presence (hatched bars) of 20 μM fumonisin B 1 . Fumonisin B 1 was added at the start of the infection (2 h p.i.), and radiolabelled serine was added at 20 h p.i. The cells were harvested 5 h later. Results are expressed as means ± standard deviations of results from three separate experiments.

    Techniques Used: Infection

    SM composition of mock-infected mouse L cells (MI), C. trachomatis -infected mouse L cells at 40 h p.i. (L2), and highly purified EBs prepared from infected mouse L cells at 40 h p.i. (EB) grown in the absence (open bars) or presence (hatched bars) of 20 μM fumonisin B 1 (A), 5 μg of brefeldin A ml −1 (B), or 0.05 U of sphingomyelinase ml −1 (C). Phospholipids were quantitated by measuring the phosphorous associated with a given phospholipid. Results for SM are expressed as percentages of the total phospholipid phosphorous. Results are averages of results from two separate experiments; duplicate results varied by less than 10%.
    Figure Legend Snippet: SM composition of mock-infected mouse L cells (MI), C. trachomatis -infected mouse L cells at 40 h p.i. (L2), and highly purified EBs prepared from infected mouse L cells at 40 h p.i. (EB) grown in the absence (open bars) or presence (hatched bars) of 20 μM fumonisin B 1 (A), 5 μg of brefeldin A ml −1 (B), or 0.05 U of sphingomyelinase ml −1 (C). Phospholipids were quantitated by measuring the phosphorous associated with a given phospholipid. Results for SM are expressed as percentages of the total phospholipid phosphorous. Results are averages of results from two separate experiments; duplicate results varied by less than 10%.

    Techniques Used: Infection, Purification

    29) Product Images from "Ceramide/Long-Chain Base Phosphate Rheostat in Saccharomyces cerevisiae: Regulation of Ceramide Synthesis by Elo3p and Cka2p"

    Article Title: Ceramide/Long-Chain Base Phosphate Rheostat in Saccharomyces cerevisiae: Regulation of Ceramide Synthesis by Elo3p and Cka2p

    Journal: Eukaryotic Cell

    doi: 10.1128/EC.2.2.284-294.2003

    Acyl-CoA-dependent ceramide synthase has a strong preference for longer-chain fatty acyls. Ceramide synthase activity was measured by the incubation of crude membranes with [ 3 H]DHS in the presence of acyl-CoA as described in Materials and Methods. (A) The reaction was linear for at least 30 min. (B) The formation of the product that comigrated with the ceramide standard, indicated by a broken circle, upon silica gel TLC was dependent on the addition of acyl-CoA and was inhibited by 10 μM australifungin and 100 μM fumonisin B 1 . (C) Crude membranes from the screening strain Y388-2 were assayed for ceramide formation after incubation with 10 μM acyl-CoAs containing saturated fatty acyls of various lengths.
    Figure Legend Snippet: Acyl-CoA-dependent ceramide synthase has a strong preference for longer-chain fatty acyls. Ceramide synthase activity was measured by the incubation of crude membranes with [ 3 H]DHS in the presence of acyl-CoA as described in Materials and Methods. (A) The reaction was linear for at least 30 min. (B) The formation of the product that comigrated with the ceramide standard, indicated by a broken circle, upon silica gel TLC was dependent on the addition of acyl-CoA and was inhibited by 10 μM australifungin and 100 μM fumonisin B 1 . (C) Crude membranes from the screening strain Y388-2 were assayed for ceramide formation after incubation with 10 μM acyl-CoAs containing saturated fatty acyls of various lengths.

    Techniques Used: Activity Assay, Incubation, Thin Layer Chromatography

    Null elo3 and cka2 mutants are supersensitive to ceramide synthase inhibitors. Cells of various deletion mutants were exposed to australifungin (A) or fumonisin B 1 (B) in the broth microdilution assay, and the growth was measured after 48 h of incubation. WT, wild type.
    Figure Legend Snippet: Null elo3 and cka2 mutants are supersensitive to ceramide synthase inhibitors. Cells of various deletion mutants were exposed to australifungin (A) or fumonisin B 1 (B) in the broth microdilution assay, and the growth was measured after 48 h of incubation. WT, wild type.

    Techniques Used: Microdilution Assay, Incubation

    Metabolism of sphingoid LCBs in S. cerevisiae. The LCBs DHS and PHS are either N acylated by ceramide synthase or phosphorylated by Lcb4 (major) and Lcb5 (minor) kinases. LCBPs can be hydrolyzed by Dpl1 lyase, forming ethanolamine-1-phosphate and palmitaldehyde, or they can be dephosphorylated by Lcb3 (DHS-1-P selective) and Ysr3 (PHS-1-P selective) lipid phosphatases. C 26 -CoA, a preferred substrate for ceramide synthase, is formed during the elongation cycle that requires the ELO3 gene. Ceramide synthase activity is sensitive to the natural product inhibitors australifungin and fumonisin B 1. As demonstrated by the results presented here, the activity of the α′ isoform of CK2 kinase encoded by CKA2 is necessary for the development of full ceramide synthase activity. MIPC, mannosyl-IPC; M(IP) 2 C, inositolphosphoryl-mannosyl-IPC.
    Figure Legend Snippet: Metabolism of sphingoid LCBs in S. cerevisiae. The LCBs DHS and PHS are either N acylated by ceramide synthase or phosphorylated by Lcb4 (major) and Lcb5 (minor) kinases. LCBPs can be hydrolyzed by Dpl1 lyase, forming ethanolamine-1-phosphate and palmitaldehyde, or they can be dephosphorylated by Lcb3 (DHS-1-P selective) and Ysr3 (PHS-1-P selective) lipid phosphatases. C 26 -CoA, a preferred substrate for ceramide synthase, is formed during the elongation cycle that requires the ELO3 gene. Ceramide synthase activity is sensitive to the natural product inhibitors australifungin and fumonisin B 1. As demonstrated by the results presented here, the activity of the α′ isoform of CK2 kinase encoded by CKA2 is necessary for the development of full ceramide synthase activity. MIPC, mannosyl-IPC; M(IP) 2 C, inositolphosphoryl-mannosyl-IPC.

    Techniques Used: Activity Assay

    30) Product Images from "Segregation of Fluorescent Membrane Lipids into Distinct Micrometric Domains: Evidence for Phase Compartmentation of Natural Lipids?"

    Article Title: Segregation of Fluorescent Membrane Lipids into Distinct Micrometric Domains: Evidence for Phase Compartmentation of Natural Lipids?

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0017021

    BODIPY-PC and -SM enriched micrometric domains show differential sensitivity to endogenous GSL and SM depletion. CHO cells were either kept untreated (a,e; CTL); selectively depleted for GSLs with the GlcCer synthase inhibitor, D-PDMP (b,f) or sphingomyelin with sphingomyelinase (SMase; c,g); or depleted of both using the upstream, dihydroceramide synthase inhibitor, fumonisin B1 (FB1; d,h), then surface-labelled with BODIPY-PC (a-d) or -SM (e-h), washed and immediately examined by confocal microscopy at 10°C. All images are bottom confocal sections recorded at the same laser power and magnification (scale bars, 2 µm). For each panel, intensity profiles along paths indicated by orange lines on confocal images are shown at right (a′-h′), by reference to baseline homogenous labelling (∼50 a.u.; horizontal dotted lines). Notice in control cells similar well-defined patches for the PC and the SM analogs (a,e) with individual sharp peaks (arrowheads #1-4). Most BODIPY-PC micrometric patches/peaks resist GSL depletion by D-PDMP (b) or SM depletion by SMase (c). In contrast, essentially all well-defined micrometric BODIPY-SM patches vanish upon either D-PDMP (f) or SMase (g). For similar properties between BODIPY-PC and -L- t -LacCer, see Fig. S4 .
    Figure Legend Snippet: BODIPY-PC and -SM enriched micrometric domains show differential sensitivity to endogenous GSL and SM depletion. CHO cells were either kept untreated (a,e; CTL); selectively depleted for GSLs with the GlcCer synthase inhibitor, D-PDMP (b,f) or sphingomyelin with sphingomyelinase (SMase; c,g); or depleted of both using the upstream, dihydroceramide synthase inhibitor, fumonisin B1 (FB1; d,h), then surface-labelled with BODIPY-PC (a-d) or -SM (e-h), washed and immediately examined by confocal microscopy at 10°C. All images are bottom confocal sections recorded at the same laser power and magnification (scale bars, 2 µm). For each panel, intensity profiles along paths indicated by orange lines on confocal images are shown at right (a′-h′), by reference to baseline homogenous labelling (∼50 a.u.; horizontal dotted lines). Notice in control cells similar well-defined patches for the PC and the SM analogs (a,e) with individual sharp peaks (arrowheads #1-4). Most BODIPY-PC micrometric patches/peaks resist GSL depletion by D-PDMP (b) or SM depletion by SMase (c). In contrast, essentially all well-defined micrometric BODIPY-SM patches vanish upon either D-PDMP (f) or SMase (g). For similar properties between BODIPY-PC and -L- t -LacCer, see Fig. S4 .

    Techniques Used: CTL Assay, Confocal Microscopy

    31) Product Images from "Invadopodia biogenesis is regulated by caveolin‐mediated modulation of membrane cholesterol levels"

    Article Title: Invadopodia biogenesis is regulated by caveolin‐mediated modulation of membrane cholesterol levels

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/j.1582-4934.2008.00568.x

    Inhibition of sphingolipid synthesis affects invadopodia formation. ( A ) A375MM cells were incubated on gelatin with BB94 in the presence or absence of 25 μg/ml fumonisin B1 for 72 hrs. BB94 was then washed out and cells incubated for further 3 hrs. Treated cells exhibited an 80% reduction of ECM degradation compared to control. ( B ) A375MM cells were incubated on gelatin and transfected without (mock) or with (GCS‐KD) siRNA targeting glucosylceramide synthase for 72 hrs. Western blot of lysates using antibodies against GCS and GAPDH as a loading control are shown. ( C ) Quantification of ECM degradation in GCS‐KD cells compared with mock. Error bars are standard deviations of the mean.
    Figure Legend Snippet: Inhibition of sphingolipid synthesis affects invadopodia formation. ( A ) A375MM cells were incubated on gelatin with BB94 in the presence or absence of 25 μg/ml fumonisin B1 for 72 hrs. BB94 was then washed out and cells incubated for further 3 hrs. Treated cells exhibited an 80% reduction of ECM degradation compared to control. ( B ) A375MM cells were incubated on gelatin and transfected without (mock) or with (GCS‐KD) siRNA targeting glucosylceramide synthase for 72 hrs. Western blot of lysates using antibodies against GCS and GAPDH as a loading control are shown. ( C ) Quantification of ECM degradation in GCS‐KD cells compared with mock. Error bars are standard deviations of the mean.

    Techniques Used: Inhibition, Incubation, Transfection, Western Blot

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    Article Snippet: .. Complementation of the lateral root defect of the loh1-1 loh3-1 mutant or FB1-treated seedlings was obtained with 0.1 μM NAA (Sigma-Aldrich) supplemented in solid Arabidopsis media with or without 0.5 μM FB1. .. The different transgenic lines used in this study were as follows: pPIN1 : PIN1-GFP , pPIN2 : PIN2-GFP , pPIN2 : PIN1-GFP , pAUX1 : AUX1-YFP , pLAX3 : LAX3-GFP , 35S : PIP2 : GFP and 35S : LTi6b : GFP , pABC19 : ABC19-GFP and pABC1 : ABC1-GFP , pSNX1 : SNX1-GFP , and pAXR4 : AXR4-GFP ( ).

    other:

    Article Title: Hepatic triglyceride accumulation via endoplasmic reticulum stress-induced SREBP-1 activation is regulated by ceramide synthases
    Article Snippet: Materials Agents were purchased/obtained as follows: (1) bortezomib (Biovision, Palo Alto, CA); (2) fumonisin B1, palmitate, 4-PBA (4-phenylbutyric acid), TUDCA (tauroursodeoxycholic acid), and anti-α-tubulin, anti-HA and anti-CerS2 antibodies (Sigma-Aldrich, St Louis, MO); (3) ceramides (fatty acyl lengths C16, C18, C20, C22, and C24) and C17 ceramide (d17:1 /C18:0 ) (Avanti Polar Lipid, Alabaster, AL); (4) anti-GRP78, anti-CHOP, anti-FAS, anti-phospho-eiF2α, anti-eiF2α, anti-PERK, and anti-SCD-1 antibodies (Cell Signaling Technology, Beverly, MA); (5) anti-SREBP-1, anti-phospho-PERK and anti-CerS6 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); (6) anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) antibody (Millipore, Temecula, CA); (7) anti-INSIG-1 antibody (Abcam, Cambridge, MA); (8) anti-mouse-HRP (horseradish peroxidase) and anti-rabbit-HRP antibodies (Jackson Laboratory, Bar Harbor, ME).

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    Article Snippet: Monoclonal mouse anti-β-actin, tubulin and fumonisin B1 were obtained from Sigma (St. Louis, MO).

    Purification:

    Article Title: Phytic acid decreases deoxynivalenol and fumonisin B1-induced changes on swine jejunal explants
    Article Snippet: .. 2.3 DON and FB1 mycotoxins The purified DON (MW: 296.32) and FB1 (MW: 721.83) mycotoxins were purchased from Sigma–Aldrich (St. Louis, MO, USA) and the Cayman Chemical Company (MI, USA), respectively. ..

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    Inhibition of ceramide synthesis rescues Atgl–/– macrophages from programmed cell death. ( a ) Wt, Atgl–/– and Atgl–/– macrophages treated with <t>FB1</t> (10 μ g/ml, 12 h) were plated on glass coverslips. Apoptosis was assessed after co-staining with FITC-conjugated annexin V (green) and PI (red), respectively, by fluorescence microscopy. Original magnification, × 10. Three fields of cells with ∼700 cells per field were counted for each condition. Data are expressed as the mean percentage of total cells±S.E.M. that stained with annexin and PI. *** P ≤0.001; ### P ≤0.001. ( b ) Total ceramide and ( c ) ceramide species were measured by LC/MS. ** P ≤0.01; # P
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    Inhibition of ceramide synthesis rescues Atgl–/– macrophages from programmed cell death. ( a ) Wt, Atgl–/– and Atgl–/– macrophages treated with FB1 (10 μ g/ml, 12 h) were plated on glass coverslips. Apoptosis was assessed after co-staining with FITC-conjugated annexin V (green) and PI (red), respectively, by fluorescence microscopy. Original magnification, × 10. Three fields of cells with ∼700 cells per field were counted for each condition. Data are expressed as the mean percentage of total cells±S.E.M. that stained with annexin and PI. *** P ≤0.001; ### P ≤0.001. ( b ) Total ceramide and ( c ) ceramide species were measured by LC/MS. ** P ≤0.01; # P

    Journal: Cell Death & Disease

    Article Title: C16 ceramide is crucial for triacylglycerol-induced apoptosis in macrophages

    doi: 10.1038/cddis.2012.17

    Figure Lengend Snippet: Inhibition of ceramide synthesis rescues Atgl–/– macrophages from programmed cell death. ( a ) Wt, Atgl–/– and Atgl–/– macrophages treated with FB1 (10 μ g/ml, 12 h) were plated on glass coverslips. Apoptosis was assessed after co-staining with FITC-conjugated annexin V (green) and PI (red), respectively, by fluorescence microscopy. Original magnification, × 10. Three fields of cells with ∼700 cells per field were counted for each condition. Data are expressed as the mean percentage of total cells±S.E.M. that stained with annexin and PI. *** P ≤0.001; ### P ≤0.001. ( b ) Total ceramide and ( c ) ceramide species were measured by LC/MS. ** P ≤0.01; # P

    Article Snippet: To inhibit ceramide synthesis, Atgl–/– macrophages were treated with 10 μ M FB1 (Calbiochem, Darmstadt, Germany) for 12 h.

    Techniques: Inhibition, Staining, Fluorescence, Microscopy, Liquid Chromatography with Mass Spectroscopy

    Inhibition of ceramide synthesis eliminates mitochondrial apoptosis and mitochondrial dysfunction in Atgl–/– macrophages. ( a ) BAX, BCL2 and cytochrome c protein levels in cytosolic fractions of Wt, Atgl–/– and FB1-treated Atgl–/– macrophages were assessed by western blotting. Data are presented as the ratios of the protein expressions relative to β -actin of two independent experiments±S.E.M. ( b ) NADH/ubiquinone oxidoreductase (complex I) and Noxa1 mRNA levels including normalization to hypoxanthine-guanine phosphoribosyltransferase (HPRT) were determined in Wt, Atgl–/– and FB1-treated Atgl–/– macrophages by real-time PCR. Data are expressed as mean values ( n =3–5)±S.E.M. ** P ≤0.01, *** P ≤0.001; ### P ≤0.001. ( c ) Mitochondrial morphology of Wt, Atgl–/– and FB1-treated Atgl–/– macrophages. Upper row: representative confocal laser-scanning microscopy images after green MitoTracker staining are shown. Lower row: representative electron micrographs. Scale bars: 0.2 μ m. ( d ) Size quantification of mitochondria visualized by electron microscopy. Data are expressed as mean ( n =45–48) ±S.E.M. *** P ≤0.001; ### P ≤0.001

    Journal: Cell Death & Disease

    Article Title: C16 ceramide is crucial for triacylglycerol-induced apoptosis in macrophages

    doi: 10.1038/cddis.2012.17

    Figure Lengend Snippet: Inhibition of ceramide synthesis eliminates mitochondrial apoptosis and mitochondrial dysfunction in Atgl–/– macrophages. ( a ) BAX, BCL2 and cytochrome c protein levels in cytosolic fractions of Wt, Atgl–/– and FB1-treated Atgl–/– macrophages were assessed by western blotting. Data are presented as the ratios of the protein expressions relative to β -actin of two independent experiments±S.E.M. ( b ) NADH/ubiquinone oxidoreductase (complex I) and Noxa1 mRNA levels including normalization to hypoxanthine-guanine phosphoribosyltransferase (HPRT) were determined in Wt, Atgl–/– and FB1-treated Atgl–/– macrophages by real-time PCR. Data are expressed as mean values ( n =3–5)±S.E.M. ** P ≤0.01, *** P ≤0.001; ### P ≤0.001. ( c ) Mitochondrial morphology of Wt, Atgl–/– and FB1-treated Atgl–/– macrophages. Upper row: representative confocal laser-scanning microscopy images after green MitoTracker staining are shown. Lower row: representative electron micrographs. Scale bars: 0.2 μ m. ( d ) Size quantification of mitochondria visualized by electron microscopy. Data are expressed as mean ( n =45–48) ±S.E.M. *** P ≤0.001; ### P ≤0.001

    Article Snippet: To inhibit ceramide synthesis, Atgl–/– macrophages were treated with 10 μ M FB1 (Calbiochem, Darmstadt, Germany) for 12 h.

    Techniques: Inhibition, Western Blot, Real-time Polymerase Chain Reaction, Confocal Laser Scanning Microscopy, Staining, Electron Microscopy

    Persistent ER stress after inhibiting ceramide synthesis in Atgl–/– macrophages. ( a ) mRNA expression of Grp78/BiP in Wt, Atgl–/– and FB1-treated Atgl–/– macrophages, including normalization to hypoxanthine-guanine phosphoribosyltransferase (HPRT), was determined by real-time PCR. ** P ≤0.01, *** P ≤0.001. ( b ) Cytosolic fractions of macrophages were isolated and proteins were resolved by SDS-PAGE. Protein expression was determined using specific antibodies for phosphorylated (p)eIF2 α , eIF2 α , and IRE1 α by western blotting. Data are expressed as the ratios of eIF2 α /eIF2 α and IRE1 α / β -actin of two independent experiments±S.E.M. CHOP protein expression was analyzed in whole cell lysates. The expression of β -actin was determined as loading control. ( c ) Wt, Atgl–/– and FB1-treated Atgl–/– macrophages were plated on coverslips in DMEM/10% LPDS. The fura-2 fluorescence ratio (340/380 nm) was determined in single macrophages before and after the addition of BHQ (15 mM) in the presence of 1 mM EGTA. Data are presented as mean values±S.E.M. of 300 cells per genotype of three independent experiments. ( d ) Area under the curve after the addition of BHQ was calculated. Dots represent means of 300 cells of three independent experiments±S.E.M. *** P ≤0.001; ## P ≤0.01. ( e ) Basal cytosolic Ca 2+ concentrations were calculated from the fura-2 fluorescence ratios (340/380 nm) during the initial 5 min. ** P ≤0.01, *** P ≤0.001; # P

    Journal: Cell Death & Disease

    Article Title: C16 ceramide is crucial for triacylglycerol-induced apoptosis in macrophages

    doi: 10.1038/cddis.2012.17

    Figure Lengend Snippet: Persistent ER stress after inhibiting ceramide synthesis in Atgl–/– macrophages. ( a ) mRNA expression of Grp78/BiP in Wt, Atgl–/– and FB1-treated Atgl–/– macrophages, including normalization to hypoxanthine-guanine phosphoribosyltransferase (HPRT), was determined by real-time PCR. ** P ≤0.01, *** P ≤0.001. ( b ) Cytosolic fractions of macrophages were isolated and proteins were resolved by SDS-PAGE. Protein expression was determined using specific antibodies for phosphorylated (p)eIF2 α , eIF2 α , and IRE1 α by western blotting. Data are expressed as the ratios of eIF2 α /eIF2 α and IRE1 α / β -actin of two independent experiments±S.E.M. CHOP protein expression was analyzed in whole cell lysates. The expression of β -actin was determined as loading control. ( c ) Wt, Atgl–/– and FB1-treated Atgl–/– macrophages were plated on coverslips in DMEM/10% LPDS. The fura-2 fluorescence ratio (340/380 nm) was determined in single macrophages before and after the addition of BHQ (15 mM) in the presence of 1 mM EGTA. Data are presented as mean values±S.E.M. of 300 cells per genotype of three independent experiments. ( d ) Area under the curve after the addition of BHQ was calculated. Dots represent means of 300 cells of three independent experiments±S.E.M. *** P ≤0.001; ## P ≤0.01. ( e ) Basal cytosolic Ca 2+ concentrations were calculated from the fura-2 fluorescence ratios (340/380 nm) during the initial 5 min. ** P ≤0.01, *** P ≤0.001; # P

    Article Snippet: To inhibit ceramide synthesis, Atgl–/– macrophages were treated with 10 μ M FB1 (Calbiochem, Darmstadt, Germany) for 12 h.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, SDS Page, Western Blot, Fluorescence

    In vitro ceramide synthase assay. (A) Enriched microsomal membranes derived from wild-type or lag1 Δ lac1 Δ (ΔΔ) cells were preincubated with wild-type cytosol, a mixture of cold and labeled dihydrosphingosine (0.5 μCi), an ATP-regenerating system, and in the presence of australifungin (lane 1) or fumonisin B1 (lane 2) for 15 min at 10°C. The ceramide synthase reaction was started by adding CoA and a liposomal mixture of hexacosanoic acid and phosphatidylinositol. After a further 120 min of incubation at 24°C, the reaction was stopped with chloroform/methanol 1:1 [vol/vol], and the lipids were hydrolyzed with mild base and desalted. Equal amounts were spotted onto HPTLC plates and resolved in solvent C. (B) CoA dependence. Experiments are performed at the same conditions as in A, except for omitting CoA (lanes 2, 3, and 5). Lipids were visualized and quantified with the use of tritium-sensitive screens and a Cyclone phosphorimager. As dihydroceramide and phytoceramide spots had very different signal intensities, a shorter exposure time for phytoceramide is shown. Abbreviations: AF, australifungin; DH-CER, dihydroceramide; FB1, fumonisin B1; PH-CER, phytoceramide. An unidentified lipid is denoted with an asterisk.

    Journal: Molecular Biology of the Cell

    Article Title: Lag1p and Lac1p Are Essential for the Acyl-CoA-dependent Ceramide Synthase Reaction in Saccharomyces cerevisae

    doi:

    Figure Lengend Snippet: In vitro ceramide synthase assay. (A) Enriched microsomal membranes derived from wild-type or lag1 Δ lac1 Δ (ΔΔ) cells were preincubated with wild-type cytosol, a mixture of cold and labeled dihydrosphingosine (0.5 μCi), an ATP-regenerating system, and in the presence of australifungin (lane 1) or fumonisin B1 (lane 2) for 15 min at 10°C. The ceramide synthase reaction was started by adding CoA and a liposomal mixture of hexacosanoic acid and phosphatidylinositol. After a further 120 min of incubation at 24°C, the reaction was stopped with chloroform/methanol 1:1 [vol/vol], and the lipids were hydrolyzed with mild base and desalted. Equal amounts were spotted onto HPTLC plates and resolved in solvent C. (B) CoA dependence. Experiments are performed at the same conditions as in A, except for omitting CoA (lanes 2, 3, and 5). Lipids were visualized and quantified with the use of tritium-sensitive screens and a Cyclone phosphorimager. As dihydroceramide and phytoceramide spots had very different signal intensities, a shorter exposure time for phytoceramide is shown. Abbreviations: AF, australifungin; DH-CER, dihydroceramide; FB1, fumonisin B1; PH-CER, phytoceramide. An unidentified lipid is denoted with an asterisk.

    Article Snippet: Before labeling, the cells were preincubated with 100 μM FB1 (Calbiochem, Bad Soden, Germany) for 2 h at 30°C as indicated.

    Techniques: In Vitro, Derivative Assay, Labeling, Incubation, High Performance Thin Layer Chromatography

    Effect of ceramide synthesis inhibitors on PA-induced insulin resistance. A: L6 myotubes were pretreated with 20 μM Fumonisin B1 or 500 nM Myriocin for 1 h, then incubated with 800 μM PA for 14 h. Subsequently, L6 myotubes were treated

    Journal: Journal of Lipid Research

    Article Title: Lysophosphatidylcholine as an effector of fatty acid-induced insulin resistance [S]

    doi: 10.1194/jlr.M014787

    Figure Lengend Snippet: Effect of ceramide synthesis inhibitors on PA-induced insulin resistance. A: L6 myotubes were pretreated with 20 μM Fumonisin B1 or 500 nM Myriocin for 1 h, then incubated with 800 μM PA for 14 h. Subsequently, L6 myotubes were treated

    Article Snippet: Fumonisin B1 and palmitoyl trifluoromethyl ketone (PACOCF3 ) were from Calbiochem (San Diego, CA).

    Techniques: Incubation

    WithaD induces N-SMase activation . (A) RT-PCR analysis of N-SMases, ceramide synthase, and A-SMase. K562 and MOLT-4 cells were treated with WithaD (1.5 and 0.5 μM respectively) for 0-120 min. RNA was extracted from total cell lysate and RT-PCR was performed. The band intensity was measured. This is one representative of three independent experiments. *indicates statistically significant difference ( P

    Journal: Molecular Cancer

    Article Title: Withanolide D induces apoptosis in leukemia by targeting the activation of neutral sphingomyelinase-ceramide cascade mediated by synergistic activation of c-Jun N-terminal kinase and p38 mitogen-activated protein kinase

    doi: 10.1186/1476-4598-9-239

    Figure Lengend Snippet: WithaD induces N-SMase activation . (A) RT-PCR analysis of N-SMases, ceramide synthase, and A-SMase. K562 and MOLT-4 cells were treated with WithaD (1.5 and 0.5 μM respectively) for 0-120 min. RNA was extracted from total cell lysate and RT-PCR was performed. The band intensity was measured. This is one representative of three independent experiments. *indicates statistically significant difference ( P

    Article Snippet: Cocktail protease inhibitor, inhibitors of N-SMase (GW4869), ceramide synthase (Fumonisin B1), JNK (SP600125) and p38MAPK (SB203580) were purchased from Calbiochem.

    Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction