lipid raft modifying drug methyl β cyclodextrin mβcd  (Millipore)


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    Name:
    Methyl beta cyclodextrin
    Description:
    Methyl β cyclodextrin is a heptasaccharide soluble in water and has more affinity to cholesterol due to the presence of hydrophobic core
    Catalog Number:
    c4555
    Price:
    None
    Applications:
    Methyl-β-cyclodextrin has been used:. to study the effect of cholesterol depletion, from sperm membrane on sperm's ability to undergo acrosome reaction.. to determine the effect of caveolin overexpression and its loss on pro-survival and pro-growth signaling. in conventional in vitro fertilization
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    Structured Review

    Millipore lipid raft modifying drug methyl β cyclodextrin mβcd
    Methyl beta cyclodextrin
    Methyl β cyclodextrin is a heptasaccharide soluble in water and has more affinity to cholesterol due to the presence of hydrophobic core
    https://www.bioz.com/result/lipid raft modifying drug methyl β cyclodextrin mβcd/product/Millipore
    Average 75 stars, based on 96 article reviews
    Price from $9.99 to $1999.99
    lipid raft modifying drug methyl β cyclodextrin mβcd - by Bioz Stars, 2020-02
    75/100 stars

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    Related Articles

    Transduction:

    Article Title: Analysis of Cd44-Containing Lipid Rafts
    Article Snippet: Mouse monoclonal antibody cross-reacting with mouse annexin II (A14020) and rabbit polyclonal antibody against caveolin (C13630) were from Transduction Laboratories and mouse monoclonal antibody against actin (mAb 1501) as well as rat monoclonal against ZO-1 (mAb 1520) were purchased from Chemikon. .. Reagents for cholesterol depletion included digitonin (D-1407) and methyl-β-cyclodextrin (M-β-CD, C-4555) both from Sigma-Aldrich.

    Cell Isolation:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: Immunocytochemical analyses B cells were purified from splenocytes using Pan B Cell Isolation kit (Miltenyi, 130–090–862) according to the manufacturer's protocol. .. Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis.

    Positive Control:

    Article Title: Isoprenoids increase bovine endometrial stromal cell tolerance to the cholesterol-dependent cytolysin from Trueperella pyogenes †
    Article Snippet: .. Methyl-β-cyclodextrin (0.5 mM, Sigma C4555) was used as a positive control because this cyclic oligosaccharide has an internal cavity that encapsulates hydrophobic sterols to efficiently deplete cholesterol from cells, which protects cells against cholesterol-dependent cytolysins [ , , ]. .. After 24-h treatment, the supernatants were discarded and the cells were challenged with control serum-free medium or medium containing 100 HU/ml pyolysin for 24 h, and cell viability was determined by MTT assay.

    Blocking Assay:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: .. Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis. .. The inhibitory effect of the compounds was verified with well-known fluorescent substrates for each endocytosis pathway, namely 100 μg/mL TF/transferrin (Life technologies, T13342) for CPZ, 0.5 μM BODIPY FL C5- Lactosylceramide/bovine serum albumin (Life Technologies, I34402) for filipin and 250 μg/mL dextran (Life Technologies, D1820) for methyl-β-cyclodextrin.

    Incubation:

    Article Title: Isoprenoids increase bovine endometrial stromal cell tolerance to the cholesterol-dependent cytolysin from Trueperella pyogenes †
    Article Snippet: To screen for molecules and pathways that might increase cell survival when cells were challenged with pyolysin, stromal cells were incubated for 24 h in serum-free medium containing vehicle, or molecules that target: MAPK (5 μM ERK Activation Inhibitor Peptide I (Calbiochem 328000) to inhibit MAPK3/1, 10 μM JNK inhibitor II (Calbiochem 420128) to inhibit MAPK8, and 10 μM SB 203580 (Calbiochem 559398) to inhibit MAPK14, as used previously [ ]); cell cycle (50 μM PNU112455A (Sigma SML0498) to inhibit cyclin dependent kinase (CDK)2 and CDK5, 80 μM roscovitine (Sigma R7772) to inhibit CDK1, CDK2, CDK5, and CDK7, and 3 μM butyrolactone I (Sigma B7930) to inhibit CDK1, CDK2, and CDK5 [ , ]); metabolic signaling pathways (10 μM AICAR (Cell Signaling 9944) to activate AMPK, 100 μM compound C (Calbiochem 171261) to inhibit AMPK, 2 μM Torin 1 (Cell Signaling 14379) to inhibit mTOR, 4 μM rapamycin (Cell Signaling 9904) to inhibit mTOR, 40 μM AKT inhibitor IV (Calbiochem 124038), and 2.8 μM LY294002 (Cell Signaling 9901) to inhibit phosphoinositide 3-kinases [ , , ]); or 48 h incubation with inhibitors for the cholesterol synthesis pathway (1 μM atorvastatin (Sigma PZ0001) to inhibit HMGCR [ ]; 100 μM etidronate (Sigma P5248) and 10 μM alendronate (Sigma A4978), which inhibit human FDPS with IC50 of 80 and 0.5 μM, respectively [ ]; 10 μM zaragozic acid (Sigma Z2626), which inhibits FDFT1 [ , ]). .. Methyl-β-cyclodextrin (0.5 mM, Sigma C4555) was used as a positive control because this cyclic oligosaccharide has an internal cavity that encapsulates hydrophobic sterols to efficiently deplete cholesterol from cells, which protects cells against cholesterol-dependent cytolysins [ , , ].

    Infection:

    Article Title: Requirements for CEACAMs and Cholesterol during Murine Coronavirus Cell Entry
    Article Snippet: Paragraph title: Infectivity determinations after cholesterol extractions and supplementations of cells. ... methyl -β-Cyclodextrin (MβCD) (Sigma catalog no. C-4555) or cholesterol-MβCD inclusion complexes (Sigma catalog no. C-4951) were serially diluted in serum-free DMEM (SFM), and 1-ml volumes were applied to 106 adherent cells in 10-cm2 dishes at ∼70 to 80% confluency for 30 min at 37°C ( , ).

    High Performance Liquid Chromatography:

    Article Title: Interaction of Cholesterol with Perfringolysin O: What Have We Learned from Functional Analysis?
    Article Snippet: .. Materials Phospholipids were from Avanti Polar Lipids); β-methyl-cyclodextrin (mCD) from Sigma C-4555 (mean degree of substitution 10.5–14.7), 7-dehydrocholesterol higher than 98% by HPLC Fluka, ergosterol 98% by HPLC Fluka, stigmasterol ~95% GC Fluka, β-sitosterol higher than 98%, and cholesterol from Steraloids. ..

    Concentration Assay:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: In some experiments, Alexa Fluor 488-labeled ScP140 control peptide was used at the same concentration. .. Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis.

    Amplex Red Cholesterol Assay:

    Article Title: Requirements for CEACAMs and Cholesterol during Murine Coronavirus Cell Entry
    Article Snippet: methyl -β-Cyclodextrin (MβCD) (Sigma catalog no. C-4555) or cholesterol-MβCD inclusion complexes (Sigma catalog no. C-4951) were serially diluted in serum-free DMEM (SFM), and 1-ml volumes were applied to 106 adherent cells in 10-cm2 dishes at ∼70 to 80% confluency for 30 min at 37°C ( , ). .. Total cell cholesterol contents before and after cholesterol depletions were measured from cell lysates using the Amplex red cholesterol assay from Molecular Probes, which detects the hydrogen peroxide product of a cholesterol oxidase enzyme reaction ( ).

    Cell Culture:

    Article Title: Real-Time Monitoring of Transferrin-Induced Endocytic Vesicle Formation by Mid-Infrared Surface Plasmon Resonance
    Article Snippet: Acute cholesterol depletion and enrichment of cells was achieved by treating the melanoma cells with methyl- β -cyclodextrin (m β CD, C-4555; Sigma, St. Louis, MO) or m β CD loaded with cholesterol (m β CD-chol), as described previously ( ). .. Cholesterol levels were determined biochemically on cells cultured at ∼70% confluence, using the Infinity cholesterol reagent kit (402-20; Sigma).

    Article Title: Isoprenoids increase bovine endometrial stromal cell tolerance to the cholesterol-dependent cytolysin from Trueperella pyogenes †
    Article Snippet: To avoid confounding cell-extrinsic factors in serum that might affect intracellular signaling pathways and cholesterol homeostasis, cells were then cultured in serum-free culture medium and treatments (see below) were applied in serum-free culture medium comprising RPMI-1640 with 50 IU/ml penicillin, 50 μg/ml streptomycin, and 2.5 μg/ml amphotericin B, as described previously [ , ]. .. Methyl-β-cyclodextrin (0.5 mM, Sigma C4555) was used as a positive control because this cyclic oligosaccharide has an internal cavity that encapsulates hydrophobic sterols to efficiently deplete cholesterol from cells, which protects cells against cholesterol-dependent cytolysins [ , , ].

    Indirect Immunoperoxidase Assay:

    Article Title: Exosomes Exploit the Virus Entry Machinery and Pathway To Transmit Alpha Interferon-Induced Antiviral Activity
    Article Snippet: .. Chemical inhibitors, including dynasore (D7693), MβCD (C4555), EIPA (A3085), IPA-3 (I2285), and rottlerin (R5648), were from Sigma-Aldrich. .. Filipin III (70440) was purchased from Cayman Chemical (Ann Arbor, MI).

    Inhibition:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: For endocytosis inhibition studies, total splenocytes or purified B cells were pretreated with endocytosis inhibitors for 30 min at 37°C, and then loaded with Alexa Fluor 488-labeled P140 peptide at 10 μM for 30 min at 37°C. .. Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis.

    Injection:

    Article Title: Roles of Specific Membrane Lipid Domains in EGF Receptor Activation and Cell Adhesion Molecule Stabilization in a Developing Olfactory System
    Article Snippet: .. In situ depletion of membrane sterols with methyl-β-cyclodextrin (MβCD) In females at early stage 2 or 3, insect saline alone (controls) or insect saline with MβCD (Sigma, #C-4555, 150 mg/ml) was injected into the headspace anterior to the brain. ..

    Recombinant:

    Article Title: Disruption of Lipid Raft Function Increases Expression and Secretion of Monocyte Chemoattractant Protein-1 in 3T3-L1 Adipocytes
    Article Snippet: Chemicals and Reagents MβCD (#C4555), water-soluble cholesterol (WSCL, #C4951), filipin complex (#F9765), BMS-345541 (#B9935), SP600125 (#S5567), and SB203580 (#S8307) were purchased from Sigma Chemical (St. Louis, MO). .. Recombinant murine TNFα (No. 410-MT) was from R & D Systems (Minneapolis, MN).

    Article Title: PGE2 mediates EGFR internalization and nuclear translocation via caveolin endocytosis promoting its transcriptional activity and proliferation in human NSCLC cells
    Article Snippet: Chemicals and reagents Recombinant human EGF (#AF-100-15) was purchased from PeproTech (Rocky Hill, NJ, USA). .. PGE2 (#P0409), Dynasore (#D7693), 5-N-Ethyl-N-isopropyl amiloride (EIPA) (#A3085) and Metil beta cyclodextrin (#C4555,) were purchased from Sigma Aldrich.

    Immunofluorescence:

    Article Title: Analysis of Cd44-Containing Lipid Rafts
    Article Snippet: Secondary antibodies used for immunofluorescence were from Jackson ImmunoResearch Laboratories: Cy3-goat anti–rat (712-165-150), Cy3-goat anti–mouse (715-160-150), FITC-goat anti–mouse (115-095-100); and from Molecular Probes: Alexa488-goat anti–rabbit (A-11008), Alexa488-goat anti–rat (A-11006) and Alexa488-goat anti–mouse (A-11001). .. Reagents for cholesterol depletion included digitonin (D-1407) and methyl-β-cyclodextrin (M-β-CD, C-4555) both from Sigma-Aldrich.

    MTT Assay:

    Article Title: Isoprenoids increase bovine endometrial stromal cell tolerance to the cholesterol-dependent cytolysin from Trueperella pyogenes †
    Article Snippet: Methyl-β-cyclodextrin (0.5 mM, Sigma C4555) was used as a positive control because this cyclic oligosaccharide has an internal cavity that encapsulates hydrophobic sterols to efficiently deplete cholesterol from cells, which protects cells against cholesterol-dependent cytolysins [ , , ]. .. After 24-h treatment, the supernatants were discarded and the cells were challenged with control serum-free medium or medium containing 100 HU/ml pyolysin for 24 h, and cell viability was determined by MTT assay.

    Microscopy:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis. .. Images were acquired with a Zeiss Axiovert LSM700 confocal microscope (Jena, Germany) with a 63× oil immersion objective.

    Purification:

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: For endocytosis inhibition studies, total splenocytes or purified B cells were pretreated with endocytosis inhibitors for 30 min at 37°C, and then loaded with Alexa Fluor 488-labeled P140 peptide at 10 μM for 30 min at 37°C. .. Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis.

    In Situ:

    Article Title: Roles of Specific Membrane Lipid Domains in EGF Receptor Activation and Cell Adhesion Molecule Stabilization in a Developing Olfactory System
    Article Snippet: .. In situ depletion of membrane sterols with methyl-β-cyclodextrin (MβCD) In females at early stage 2 or 3, insect saline alone (controls) or insect saline with MβCD (Sigma, #C-4555, 150 mg/ml) was injected into the headspace anterior to the brain. ..

    Activation Assay:

    Article Title: Isoprenoids increase bovine endometrial stromal cell tolerance to the cholesterol-dependent cytolysin from Trueperella pyogenes †
    Article Snippet: To screen for molecules and pathways that might increase cell survival when cells were challenged with pyolysin, stromal cells were incubated for 24 h in serum-free medium containing vehicle, or molecules that target: MAPK (5 μM ERK Activation Inhibitor Peptide I (Calbiochem 328000) to inhibit MAPK3/1, 10 μM JNK inhibitor II (Calbiochem 420128) to inhibit MAPK8, and 10 μM SB 203580 (Calbiochem 559398) to inhibit MAPK14, as used previously [ ]); cell cycle (50 μM PNU112455A (Sigma SML0498) to inhibit cyclin dependent kinase (CDK)2 and CDK5, 80 μM roscovitine (Sigma R7772) to inhibit CDK1, CDK2, CDK5, and CDK7, and 3 μM butyrolactone I (Sigma B7930) to inhibit CDK1, CDK2, and CDK5 [ , ]); metabolic signaling pathways (10 μM AICAR (Cell Signaling 9944) to activate AMPK, 100 μM compound C (Calbiochem 171261) to inhibit AMPK, 2 μM Torin 1 (Cell Signaling 14379) to inhibit mTOR, 4 μM rapamycin (Cell Signaling 9904) to inhibit mTOR, 40 μM AKT inhibitor IV (Calbiochem 124038), and 2.8 μM LY294002 (Cell Signaling 9901) to inhibit phosphoinositide 3-kinases [ , , ]); or 48 h incubation with inhibitors for the cholesterol synthesis pathway (1 μM atorvastatin (Sigma PZ0001) to inhibit HMGCR [ ]; 100 μM etidronate (Sigma P5248) and 10 μM alendronate (Sigma A4978), which inhibit human FDPS with IC50 of 80 and 0.5 μM, respectively [ ]; 10 μM zaragozic acid (Sigma Z2626), which inhibits FDFT1 [ , ]). .. Methyl-β-cyclodextrin (0.5 mM, Sigma C4555) was used as a positive control because this cyclic oligosaccharide has an internal cavity that encapsulates hydrophobic sterols to efficiently deplete cholesterol from cells, which protects cells against cholesterol-dependent cytolysins [ , , ].

    FLAG-tag:

    Article Title: Deletion of OSBPL2 in auditory cells increases cholesterol biosynthesis and drives reactive oxygen species production by inhibiting AMPK activity
    Article Snippet: Antibodies and reagents Antibodies for adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK/Ampk) (#2793), Flag-Tag (#2368), His-Tag (#12698), GST-Tag (#2624), GAPDH (#5174) and phosphospecific antibodies for AMPK/Ampk -pThr172 (#2535) were purchased from Cell Signaling Technology (Danvers, MA, USA). .. Cholesterol (#C8667) and methyl-β-cyclodextrin (#C4555) were purchased from Sigma-ldrich (St. Louis, MO, USA).

    Staining:

    Article Title: The tumour suppressor OPCML promotes AXL inactivation by the phosphatase PTPRG in ovarian cancer
    Article Snippet: Methyl‐β‐cyclodextrin (MBCD, #C4555, Sigma‐Aldrich) was used to deplete cholesterol. .. The cells were stained with 0.05 mg/ml filipin (#C4767, Sigma‐Aldrich) diluted in PBS for 1 h at room temperature and rinsed thrice with PBS.

    Article Title: Modulation of deregulated chaperone-mediated autophagy by a phosphopeptide
    Article Snippet: Ten μg/mL CPZ (Sigma-Aldrich, C-8138) was used to inhibit clathrin-mediated endocytosis, 5 μg/mL filipin (Sigma-Aldrich, F-4767) to disrupt caveolae-dependent endocytosis, and 5 mM methyl-β-cyclodextrin (Sigma-Aldrich, C-4555) to block macropinocytosis. .. Cells were then permeabilized with 0.05% (v/v) of Triton X-100 (Pharmacia Biotech, 17–1315–01), stained with 4′,6-diamidino-2-phenylindole (DAPI) (Molecular Probes, D1306), and finally mounted on glass slides with fluorescent mounting medium (DAKO, S302380).

    other:

    Article Title: Cholesterol Protects Against Acute Stress-Induced T-Tubule Remodeling in Mouse Ventricular Myocytes
    Article Snippet: Methyl-β-cyclodextrin (MβCD; C4555), Water Soluble Cholesterol (WSC; C4951), Filipin (F9765) and all other chemicals and reagents were purchased from Sigma, St. Louis, MO, United States.

    Article Title: Follicular Stimulating Hormone Accelerates Atherogenesis by Increasing Endothelial VCAM-1 Expression
    Article Snippet: Materials FSH (#F4021), 17β-estradiol (E2, #E2758), TNF-α(#T0157), pertussis toxin (PTX, #3097), β-methyl cyclodextrin (β-MCD, #C-4555), and KU0063794 (#SML0382) were purchased from Sigma Aldrich (St. Louis, MO, USA).

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  • 95
    Millipore cholesterol mβcd
    Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol <t>methyl-β-cyclodextrin</t> (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P
    Cholesterol Mβcd, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cholesterol mβcd/product/Millipore
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    cholesterol mβcd - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    77
    Millipore meβcd
    Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or <t>MeβCD</t>
    Meβcd, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/meβcd/product/Millipore
    Average 77 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    meβcd - by Bioz Stars, 2020-02
    77/100 stars
      Buy from Supplier

    79
    Millipore mβcd powder
    Recovery of dysregulated protein signature in NPC1 fibroblasts upon <t>MβCD</t> treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.
    Mβcd Powder, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mβcd powder/product/Millipore
    Average 79 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mβcd powder - by Bioz Stars, 2020-02
    79/100 stars
      Buy from Supplier

    Image Search Results


    Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol methyl-β-cyclodextrin (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P

    Journal: JCI Insight

    Article Title: AIBP augments cholesterol efflux from alveolar macrophages to surfactant and reduces acute lung inflammation

    doi: 10.1172/jci.insight.120519

    Figure Lengend Snippet: Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol methyl-β-cyclodextrin (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P

    Article Snippet: In some experiments, cells were incubated for 1 hour with or without 50 μg/ml cholesterol-MβCD (MilliporeSigma, C4951).

    Techniques:

    Macrophages release particles enriched in cholesterol. ( A ) Macrophages were loaded with cholesterol/MβCD. SEM and nanoSIMS images of the macrophage after a short incubation with [ 15 N]ALO-D4. The SEM image shows a lawn of particles outside the macrophage; the nanoSIMS image (scaled at two different settings) reveals binding of [ 15 N]ALO-D4 to the particles, indicating that they contain accessible cholesterol. The boxed regions are shown below at higher magnification, again showing binding of [ 15 N]ALO-D4 to macrophage-derived particles. (Scale bars, 5 μm.) The bar graph shows 15 N/ 14 N levels for the cell body and particles of two cells (60 particles were quantified). The y axis starts at 0.0037, the natural abundance of 15 N. Data are shown as mean ± SD. ( B ) Macrophages were loaded with acetyl-LDL. Particles released by macrophages that had been loaded with acetyl-LDL (50 μg/mL) (yellow arrows) are visible in secondary electron (SE) and 12 C 14 N − nanoSIMS images. Composite 12 C 14 N − or secondary electron (SE) (gray) and 15 N/ 14 N ratio (red) images show binding of [ 15 N]ALO-D4 to particles. (Scale bars, 2 μm.)

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

    Article Title: Macrophages release plasma membrane-derived particles rich in accessible cholesterol

    doi: 10.1073/pnas.1810724115

    Figure Lengend Snippet: Macrophages release particles enriched in cholesterol. ( A ) Macrophages were loaded with cholesterol/MβCD. SEM and nanoSIMS images of the macrophage after a short incubation with [ 15 N]ALO-D4. The SEM image shows a lawn of particles outside the macrophage; the nanoSIMS image (scaled at two different settings) reveals binding of [ 15 N]ALO-D4 to the particles, indicating that they contain accessible cholesterol. The boxed regions are shown below at higher magnification, again showing binding of [ 15 N]ALO-D4 to macrophage-derived particles. (Scale bars, 5 μm.) The bar graph shows 15 N/ 14 N levels for the cell body and particles of two cells (60 particles were quantified). The y axis starts at 0.0037, the natural abundance of 15 N. Data are shown as mean ± SD. ( B ) Macrophages were loaded with acetyl-LDL. Particles released by macrophages that had been loaded with acetyl-LDL (50 μg/mL) (yellow arrows) are visible in secondary electron (SE) and 12 C 14 N − nanoSIMS images. Composite 12 C 14 N − or secondary electron (SE) (gray) and 15 N/ 14 N ratio (red) images show binding of [ 15 N]ALO-D4 to particles. (Scale bars, 2 μm.)

    Article Snippet: Macrophages were loaded with 20 μL/mL of [13 C]cholesterol/MβCD (final cholesterol concentration, 50 μM) in macrophage medium containing 0.1% LPDS, 50 μM mevastatin (Calbiochem), and 50 μM mevalonolactone (Sigma) for 24 h at 37 °C.

    Techniques: Incubation, Binding Assay, Derivative Assay

    Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Progesterone- and MeβCD-stimulated increases in oocyte 39-kDa Mos are inhibited by cycloheximide

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Progesterone- and MeβCD-stimulated increases in oocyte 39-kDa Mos are inhibited by cycloheximide

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    The steroid synthesis inhibitor, aminoglutethimide, does not block oocyte maturation stimulated MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: The steroid synthesis inhibitor, aminoglutethimide, does not block oocyte maturation stimulated MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques: Blocking Assay

    Follicle cells are not required for phosphorylation of oocyte MAPK or GVBD induced by MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Follicle cells are not required for phosphorylation of oocyte MAPK or GVBD induced by MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Cycloheximide blocks MAPK phosphorylation and GVBD in response to MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Cycloheximide blocks MAPK phosphorylation and GVBD in response to MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Recovery of dysregulated protein signature in NPC1 fibroblasts upon MβCD treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Recovery of dysregulated protein signature in NPC1 fibroblasts upon MβCD treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques:

    Effects of different sources of MβCDs on reducing lysosome size in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days, after which LysoTracker ® staining was performed. (A) Images of LysoTracker staining on NPC1 fibroblasts. Treatment with 300 and 11 μM of MβCD-3 significantly reduced the cholesterol accumulation in NPC1 patient skin fibroblasts, while there were no significant effects observed from the other two batches of MβCDs (MβCD-1 and MβCD-2). LysoTracker red stains cellular acidic compartments to visualize enlarged lysosomes, and Hoechst ( blue ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced the lysosome size in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts. MβCD-3 showed concentration-dependent impact on lysosome size of NPC1 fibroblasts, while there were no significant effects observed from the other two batches of MβCDs.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effects of different sources of MβCDs on reducing lysosome size in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days, after which LysoTracker ® staining was performed. (A) Images of LysoTracker staining on NPC1 fibroblasts. Treatment with 300 and 11 μM of MβCD-3 significantly reduced the cholesterol accumulation in NPC1 patient skin fibroblasts, while there were no significant effects observed from the other two batches of MβCDs (MβCD-1 and MβCD-2). LysoTracker red stains cellular acidic compartments to visualize enlarged lysosomes, and Hoechst ( blue ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced the lysosome size in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts. MβCD-3 showed concentration-dependent impact on lysosome size of NPC1 fibroblasts, while there were no significant effects observed from the other two batches of MβCDs.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Staining, Concentration Assay

    Mass spectrometry on MβCD. Mass spectra of MβCD-1, -2, and -3 show cluster signals of sodium adduct ions of 6–12 different mixtures of methyl-substituted β-cyclodextrin molecules. Methylation number can be easily determined by an inclement of mass unit (m/z) 14 as shown. The mass spectrometry peak heights are proportional to molecular distributions (or amounts) of various methyl-substituted β-cyclodextrins (summarized in Table 1 ). Abundance of lower number of methyl substitution of MβCD is shown in (A) (MβCD-1), while middle substitution is in (B) (MβCD-2) and higher substitution is in (C) (MβCD-3). 10-Me, 10 methylation to β-cyclodextrin molecule.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Mass spectrometry on MβCD. Mass spectra of MβCD-1, -2, and -3 show cluster signals of sodium adduct ions of 6–12 different mixtures of methyl-substituted β-cyclodextrin molecules. Methylation number can be easily determined by an inclement of mass unit (m/z) 14 as shown. The mass spectrometry peak heights are proportional to molecular distributions (or amounts) of various methyl-substituted β-cyclodextrins (summarized in Table 1 ). Abundance of lower number of methyl substitution of MβCD is shown in (A) (MβCD-1), while middle substitution is in (B) (MβCD-2) and higher substitution is in (C) (MβCD-3). 10-Me, 10 methylation to β-cyclodextrin molecule.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Mass Spectrometry, Methylation

    Chemical structure of β-cyclodextrin. (A) Chemical representation of methyl-β-cyclodextrin (MβCD), which comprises seven glucopyranose units. (B) Three-dimensional representation of the toroid structure of cyclodextrin consisting of a hydrophilic exterior and hydrophobic interior.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Chemical structure of β-cyclodextrin. (A) Chemical representation of methyl-β-cyclodextrin (MβCD), which comprises seven glucopyranose units. (B) Three-dimensional representation of the toroid structure of cyclodextrin consisting of a hydrophilic exterior and hydrophobic interior.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques:

    Effects of different sources of MβCDs on reducing cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days; filipin staining was then performed. (A) Images of filipin staining on NPC1 fibroblasts. Treatment with 300, 11 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 patient skin fibroblasts, while the other two batches of MβCDs (MβCD-1 and MβCD-2) showed much weaker effects on cholesterol accumulation in NPC1 patient fibroblasts. Filipin ( green ) stains the intracellular cholesterol-laden domains, and EthD-1 ( red ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced cholesterol accumulation in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on cholesterol accumulation in NPC1 patient fibroblasts. EthD-1, ethidium homodimer; NPC1, Niemann-Pick disease type C1; WT, wild-type.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effects of different sources of MβCDs on reducing cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days; filipin staining was then performed. (A) Images of filipin staining on NPC1 fibroblasts. Treatment with 300, 11 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 patient skin fibroblasts, while the other two batches of MβCDs (MβCD-1 and MβCD-2) showed much weaker effects on cholesterol accumulation in NPC1 patient fibroblasts. Filipin ( green ) stains the intracellular cholesterol-laden domains, and EthD-1 ( red ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced cholesterol accumulation in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on cholesterol accumulation in NPC1 patient fibroblasts. EthD-1, ethidium homodimer; NPC1, Niemann-Pick disease type C1; WT, wild-type.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Staining, Ethidium Homodimer Assay

    Effect of different sources of MβCDs on cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were treated with MβCD (0.4–300 μM) for 4 days, followed by an Amplex ® Red cholesterol assay. (A) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts on cholesterol accumulation in NPC1 fibroblasts. MβCD-3 showed concentration-dependent manner on cholesterol accumulation in NPC1 fibroblasts GM03123, while there were much weaker effects observed with the other two batches of MβCDs. The maximum inhibitory effect of MβCD-3 is about 46.0% compared with MβCD-1, 15.2%, and MβCD-2, 19.5%. (B) Treatment with 300 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 fibroblasts compared with the other two sources of MβCDs. (C) Cytotoxicity (ATPlite assay) of different sources of MβCDs on NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were untreated or treated with MβCD (0.4–300 μM) for 4 days and ATPlite assay was performed to evaluate the cytotoxic effects of different sources of MβCDs on the fibroblasts. There were no significant cytotoxic effects observed within the test range of 0.4–300 μM of MβCD and the cell viability level was generally above 90% after 4 days of treatment.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effect of different sources of MβCDs on cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were treated with MβCD (0.4–300 μM) for 4 days, followed by an Amplex ® Red cholesterol assay. (A) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts on cholesterol accumulation in NPC1 fibroblasts. MβCD-3 showed concentration-dependent manner on cholesterol accumulation in NPC1 fibroblasts GM03123, while there were much weaker effects observed with the other two batches of MβCDs. The maximum inhibitory effect of MβCD-3 is about 46.0% compared with MβCD-1, 15.2%, and MβCD-2, 19.5%. (B) Treatment with 300 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 fibroblasts compared with the other two sources of MβCDs. (C) Cytotoxicity (ATPlite assay) of different sources of MβCDs on NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were untreated or treated with MβCD (0.4–300 μM) for 4 days and ATPlite assay was performed to evaluate the cytotoxic effects of different sources of MβCDs on the fibroblasts. There were no significant cytotoxic effects observed within the test range of 0.4–300 μM of MβCD and the cell viability level was generally above 90% after 4 days of treatment.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Amplex Red Cholesterol Assay, Concentration Assay