c2c12  (Roche)


Bioz Verified Symbol Roche is a verified supplier
Bioz Manufacturer Symbol Roche manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92

    Structured Review

    Roche c2c12
    BCKAs impair insulin signaling in <t>C2C12</t> cells. (a-c) Differentiated C2C12 cells were pre-incubated with either 2% BSA or 2% BSA conjugated with 0.4mM palmitate for 16 hr followed by either a) 5mM ketoleucine, b) ketovaline or c) ketoisoleucine treatment for 30 mins. Myotubes were subjected to 100 nM insulin for 15 mins in presence or absence of individual BCKAs. Immunoblot and densitometric analysis of total and phosphorylated AKT Ser 473 and Thr 309, total and phosphorylated IRS Tyr 612. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p
    C2c12, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 4401 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/c2c12/product/Roche
    Average 92 stars, based on 4401 article reviews
    Price from $9.99 to $1999.99
    c2c12 - by Bioz Stars, 2020-09
    92/100 stars

    Images

    1) Product Images from "Silencing branched-chain ketoacid dehydrogenase or treatment with branched-chain ketoacids ex vivo inhibits muscle insulin signaling"

    Article Title: Silencing branched-chain ketoacid dehydrogenase or treatment with branched-chain ketoacids ex vivo inhibits muscle insulin signaling

    Journal: bioRxiv

    doi: 10.1101/2020.02.21.960153

    BCKAs impair insulin signaling in C2C12 cells. (a-c) Differentiated C2C12 cells were pre-incubated with either 2% BSA or 2% BSA conjugated with 0.4mM palmitate for 16 hr followed by either a) 5mM ketoleucine, b) ketovaline or c) ketoisoleucine treatment for 30 mins. Myotubes were subjected to 100 nM insulin for 15 mins in presence or absence of individual BCKAs. Immunoblot and densitometric analysis of total and phosphorylated AKT Ser 473 and Thr 309, total and phosphorylated IRS Tyr 612. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p
    Figure Legend Snippet: BCKAs impair insulin signaling in C2C12 cells. (a-c) Differentiated C2C12 cells were pre-incubated with either 2% BSA or 2% BSA conjugated with 0.4mM palmitate for 16 hr followed by either a) 5mM ketoleucine, b) ketovaline or c) ketoisoleucine treatment for 30 mins. Myotubes were subjected to 100 nM insulin for 15 mins in presence or absence of individual BCKAs. Immunoblot and densitometric analysis of total and phosphorylated AKT Ser 473 and Thr 309, total and phosphorylated IRS Tyr 612. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p

    Techniques Used: Incubation

    BCKAs activate mTORC1 and downstream protein translation signaling in skeletal muscle. a) Schematic of mTORC1 signaling and its downstream protein translational targets. b) C2C12 myotubes were treated with 5mM ketoleucine, ketoisoleucine and ketovaline for 45 mins. b-c) Immunoblot analysis and densitometric quantification of total and phosphorylated mTORC1 Ser 2448, p70S6K Thr 389, eEF2 Thr 56, eIFG Ser 1108 in C2C12 myotubes transduced with Ad-CMV-shGFP or Ad-CMV-shGFP-r/mBCKDHA for 48 hr followed by stimulation with 100nM insulin for 15 mins. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p
    Figure Legend Snippet: BCKAs activate mTORC1 and downstream protein translation signaling in skeletal muscle. a) Schematic of mTORC1 signaling and its downstream protein translational targets. b) C2C12 myotubes were treated with 5mM ketoleucine, ketoisoleucine and ketovaline for 45 mins. b-c) Immunoblot analysis and densitometric quantification of total and phosphorylated mTORC1 Ser 2448, p70S6K Thr 389, eEF2 Thr 56, eIFG Ser 1108 in C2C12 myotubes transduced with Ad-CMV-shGFP or Ad-CMV-shGFP-r/mBCKDHA for 48 hr followed by stimulation with 100nM insulin for 15 mins. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p

    Techniques Used: Transduction

    Okadaic acid rescue BCKAs induced insulin signaling impairment. a) Outline of the treatment scheme used to test the effect of okadaic acid on ketoleucine mediated insulin signaling. b) PP2A activity expressed as percent mean control and c) immunoblot and densitometric analysis of total to phosphorylated AKT Ser 473 and total to phosphorylated AKT2 Ser 474 in differentiated C2C12 myotubes pretreated with 250nM okadaic acid (OA) for 30 mins followed by 5mM ketoleucine for 30 mins and 100nM insulin stimulation for 15 mins. No insulin-treated cells were employed as controls. Other relevant groups are provided as an illustration in (a). Statistical analysis was performed using a one-way ANOVA followed by a Tukey’s multiple comparison test; *p
    Figure Legend Snippet: Okadaic acid rescue BCKAs induced insulin signaling impairment. a) Outline of the treatment scheme used to test the effect of okadaic acid on ketoleucine mediated insulin signaling. b) PP2A activity expressed as percent mean control and c) immunoblot and densitometric analysis of total to phosphorylated AKT Ser 473 and total to phosphorylated AKT2 Ser 474 in differentiated C2C12 myotubes pretreated with 250nM okadaic acid (OA) for 30 mins followed by 5mM ketoleucine for 30 mins and 100nM insulin stimulation for 15 mins. No insulin-treated cells were employed as controls. Other relevant groups are provided as an illustration in (a). Statistical analysis was performed using a one-way ANOVA followed by a Tukey’s multiple comparison test; *p

    Techniques Used: Activity Assay

    Pharmacological inhibition of BCKDK by BT2 improves muscle insulin signaling. a) A diagrammatic representation of BT2 action. b) Intracellular BCKAs by UPLC/MS-MS and c) qPCR analysis of BCAA catabolic genes Bckdha , Bckdk , Bcat2 , Hadh , Hibch and Klf15 corrected to Rer1/Rpl7 reference gene levels in C2C12 myotubes treated with 500µM and 750µM BT2 for 20 hr. d) Immunoblot analysis and densitometric quantification of phosphorylated BCKDH subunit E1 at Ser 293 in C2C12 myotubes treated with 160µM, 250µM, 320µM, 500µM and 750µM BT2 for 20 hr. e) Immunoblotting and densitometric quantification of phosphorylated AKT Ser 473 and total AKT in differentiated C2C12 cells pretreated with 320µM, 500µM and 750µM BT2 for 20 hr followed by incubation with 100nM insulin for 15 mins. (f) Immunoblot analysis and densitometric quantification of phosphorylated AKT Ser 473, Ser 474 and total AKT1 and AKT2 in C2C12 myoblasts transduced with Ad-CMV-shGFP or Ad-CMV-shGFP-r/mBCKDHA (MOI 200) for 48 hr followed by incubation with 500µM BT2 for 20 hr and stimulation with 100nM insulin for 15 mins. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p
    Figure Legend Snippet: Pharmacological inhibition of BCKDK by BT2 improves muscle insulin signaling. a) A diagrammatic representation of BT2 action. b) Intracellular BCKAs by UPLC/MS-MS and c) qPCR analysis of BCAA catabolic genes Bckdha , Bckdk , Bcat2 , Hadh , Hibch and Klf15 corrected to Rer1/Rpl7 reference gene levels in C2C12 myotubes treated with 500µM and 750µM BT2 for 20 hr. d) Immunoblot analysis and densitometric quantification of phosphorylated BCKDH subunit E1 at Ser 293 in C2C12 myotubes treated with 160µM, 250µM, 320µM, 500µM and 750µM BT2 for 20 hr. e) Immunoblotting and densitometric quantification of phosphorylated AKT Ser 473 and total AKT in differentiated C2C12 cells pretreated with 320µM, 500µM and 750µM BT2 for 20 hr followed by incubation with 100nM insulin for 15 mins. (f) Immunoblot analysis and densitometric quantification of phosphorylated AKT Ser 473, Ser 474 and total AKT1 and AKT2 in C2C12 myoblasts transduced with Ad-CMV-shGFP or Ad-CMV-shGFP-r/mBCKDHA (MOI 200) for 48 hr followed by incubation with 500µM BT2 for 20 hr and stimulation with 100nM insulin for 15 mins. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p

    Techniques Used: Inhibition, Tandem Mass Spectroscopy, Real-time Polymerase Chain Reaction, Incubation, Transduction

    Modulating intracellular BCKAs by modifying BCAA catabolic enzyme expression alters skeletal and cardiac muscle insulin signaling. a) Brief schematic of enzymes regulating BCKA catabolism. b) UPLC mass spectrometric analysis of intracellular BCKAs levels in C2C12 myotubes transduced with Ad-CMV-GFP-hBCKDK/HA (MOI 150) or Ad-CMV-mCherry-hBCKDHA/Flag (MOI 150) and their appropriate controls. The graph represents mean ± S.E.M., n=3, *p
    Figure Legend Snippet: Modulating intracellular BCKAs by modifying BCAA catabolic enzyme expression alters skeletal and cardiac muscle insulin signaling. a) Brief schematic of enzymes regulating BCKA catabolism. b) UPLC mass spectrometric analysis of intracellular BCKAs levels in C2C12 myotubes transduced with Ad-CMV-GFP-hBCKDK/HA (MOI 150) or Ad-CMV-mCherry-hBCKDHA/Flag (MOI 150) and their appropriate controls. The graph represents mean ± S.E.M., n=3, *p

    Techniques Used: Expressing, Transduction

    2) Product Images from "Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary"

    Article Title: Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.17039

    Simultaneous localization of non-methylated and methylated GATCG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Sau 3A I cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Mbo I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images are given in C, F and I . Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated GATCG was localized in the nuclei of granulosa cells (white arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.
    Figure Legend Snippet: Simultaneous localization of non-methylated and methylated GATCG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Sau 3A I cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Mbo I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images are given in C, F and I . Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated GATCG was localized in the nuclei of granulosa cells (white arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.

    Techniques Used: Methylation, Labeling, Staining

    Simultaneous localization of non-methylated and methylated CCGG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Hpa II cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Msp I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images were shown in C, F and I . A, B and C : Primary follicle . D, E and F : Secondary follicle. G, H and I : Tertiary follicle. Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated CCGG was localized in the nuclei of granulosa cells (white arrows) or oocytes (red arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.
    Figure Legend Snippet: Simultaneous localization of non-methylated and methylated CCGG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Hpa II cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Msp I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images were shown in C, F and I . A, B and C : Primary follicle . D, E and F : Secondary follicle. G, H and I : Tertiary follicle. Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated CCGG was localized in the nuclei of granulosa cells (white arrows) or oocytes (red arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.

    Techniques Used: Methylation, Labeling, Staining

    3) Product Images from "Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages"

    Article Title: Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0123293

    LOX-1 is capable of binding HOCl-modified proteins, but does not contribute to OVA-Cl uptake by BM-DC. ( A ) Binding of rLOX-1 to plate-adsorbed proteins. ( B , C ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rLOX-1 binding to plate-adsorbed OVA-Cl (B) or GA-BSA (C). ( D ) Binding of PE-conjugated anti-mouse LOX-1 mAb and control rat IgG2a to CBA BM-DC, determined by flow cytometry. ( E ) The effect of blocking goat anti-mouse LOX-1 polyclonal Ab, relative to normal goat IgG, on AF-OVA-Cl uptake by untreated and LPS-pre-treated CBA BM-DC. ( F ) LOX-1 expression on LPS-pre-treated CBA BM-DC. Results of single experiments are shown, repeated at least twice with similar results. The data were analysed by the Student’s t-test (A, E) or by ANOVA, followed by the Tukey-Kramer post-test (B, C). *, p
    Figure Legend Snippet: LOX-1 is capable of binding HOCl-modified proteins, but does not contribute to OVA-Cl uptake by BM-DC. ( A ) Binding of rLOX-1 to plate-adsorbed proteins. ( B , C ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rLOX-1 binding to plate-adsorbed OVA-Cl (B) or GA-BSA (C). ( D ) Binding of PE-conjugated anti-mouse LOX-1 mAb and control rat IgG2a to CBA BM-DC, determined by flow cytometry. ( E ) The effect of blocking goat anti-mouse LOX-1 polyclonal Ab, relative to normal goat IgG, on AF-OVA-Cl uptake by untreated and LPS-pre-treated CBA BM-DC. ( F ) LOX-1 expression on LPS-pre-treated CBA BM-DC. Results of single experiments are shown, repeated at least twice with similar results. The data were analysed by the Student’s t-test (A, E) or by ANOVA, followed by the Tukey-Kramer post-test (B, C). *, p

    Techniques Used: Binding Assay, Modification, Crocin Bleaching Assay, Flow Cytometry, Cytometry, Blocking Assay, Expressing

    Both SREC-I and RAGE bind HOCl-oxidised proteins with low affinities. ( A ) Binding of rSREC-I to proteins coated onto ELISA plates. ( B ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rSREC-I binding to plate-adsorbed AcLDL. ( C ) Binding of goat anti-mouse SREC-I Ab to untreated and LPS-pre-treated BM-DC. ( D ) Effects of polyclonal goat anti-mouse SREC-I Ab, relative to normal goat IgG, on the uptake of AF-AcLDL, AF-OVA-Cl and pHr-HSA-Cl by LPS-pre-treated, SR-A-deficient BM-DC. ( E ) Binding of rRAGE to plate-adsorbed proteins. ( F ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rRAGE binding to plate-adsorbed GA-BSA. The data were analysed by the Student’s t-test (A, D, E) or by ANOVA, followed by the Tukey-Kramer post-test (B, F). *, p
    Figure Legend Snippet: Both SREC-I and RAGE bind HOCl-oxidised proteins with low affinities. ( A ) Binding of rSREC-I to proteins coated onto ELISA plates. ( B ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rSREC-I binding to plate-adsorbed AcLDL. ( C ) Binding of goat anti-mouse SREC-I Ab to untreated and LPS-pre-treated BM-DC. ( D ) Effects of polyclonal goat anti-mouse SREC-I Ab, relative to normal goat IgG, on the uptake of AF-AcLDL, AF-OVA-Cl and pHr-HSA-Cl by LPS-pre-treated, SR-A-deficient BM-DC. ( E ) Binding of rRAGE to plate-adsorbed proteins. ( F ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rRAGE binding to plate-adsorbed GA-BSA. The data were analysed by the Student’s t-test (A, D, E) or by ANOVA, followed by the Tukey-Kramer post-test (B, F). *, p

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    4) Product Images from "Lysophosphatidic acid-induced p21Waf1 expression mediates the cytostatic response of breast and ovarian cancer cells to transforming growth factor beta"

    Article Title: Lysophosphatidic acid-induced p21Waf1 expression mediates the cytostatic response of breast and ovarian cancer cells to transforming growth factor beta

    Journal: Molecular Cancer Research

    doi: 10.1158/1541-7786.MCR-11-0340

    p21-dependent inhibition of LPA-induced cell proliferation by TGFβ. A . TGFβ inhibits LPA-afforded cell proliferation. MDA-MB-231 and Caov-3 cells in 6-well plates were incubated with LPA (10 µM) or vehicle (BSA) in the presence
    Figure Legend Snippet: p21-dependent inhibition of LPA-induced cell proliferation by TGFβ. A . TGFβ inhibits LPA-afforded cell proliferation. MDA-MB-231 and Caov-3 cells in 6-well plates were incubated with LPA (10 µM) or vehicle (BSA) in the presence

    Techniques Used: Inhibition, Multiple Displacement Amplification, Incubation

    5) Product Images from "Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary"

    Article Title: Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.17039

    Methylation level of CCGG sites in TUNEL-positive follicle cells in adjacent sections of mouse ovary. Adjacent sections of paraffin-embedded mouse ovary were used for H E staining, TUNEL, and the detection of non-methylated CCGG sites and methylated CCGG sites, as described in detail in “II. Materials and Methods”. A : H E staining. B : TUNEL staining. C : Blockade of 3'-OH ends with dideoxynucleotides by TdT. After blockade, the section was labeled with biotin-16-dUTP by TdT and the incorporated biotin was detected with HRP-anti-biotin. No signals were observed. D : Staining for non-methylated CCGG sites. After the blockade procedure described in ( C ), the section was digested with Hpa II, labeled with biotin-16-dUTP and visualized by enzyme-immunohistochemistry with HRP-anti-biotin. E : Blockade of Hpa II cutting sites with dideoxynucleotides by TdT. The section was digested with Hpa II and the cutting sites were blocked with a dideoxynucleotide mixture. Then the section was processed in a manner similar to that described in ( C ). F : Staining for methylated CCGG sites. After blockade of Hpa II cutting sites with dideoxynucleotides, the section was digested with Msp I and the cutting sites were labeled with biotin-16-dUTP and visualized with HRP-anti-biotin. Most of TUNEL-positive follicle cells were found in methylation level of CCGG sites. Bar = 100 μm.
    Figure Legend Snippet: Methylation level of CCGG sites in TUNEL-positive follicle cells in adjacent sections of mouse ovary. Adjacent sections of paraffin-embedded mouse ovary were used for H E staining, TUNEL, and the detection of non-methylated CCGG sites and methylated CCGG sites, as described in detail in “II. Materials and Methods”. A : H E staining. B : TUNEL staining. C : Blockade of 3'-OH ends with dideoxynucleotides by TdT. After blockade, the section was labeled with biotin-16-dUTP by TdT and the incorporated biotin was detected with HRP-anti-biotin. No signals were observed. D : Staining for non-methylated CCGG sites. After the blockade procedure described in ( C ), the section was digested with Hpa II, labeled with biotin-16-dUTP and visualized by enzyme-immunohistochemistry with HRP-anti-biotin. E : Blockade of Hpa II cutting sites with dideoxynucleotides by TdT. The section was digested with Hpa II and the cutting sites were blocked with a dideoxynucleotide mixture. Then the section was processed in a manner similar to that described in ( C ). F : Staining for methylated CCGG sites. After blockade of Hpa II cutting sites with dideoxynucleotides, the section was digested with Msp I and the cutting sites were labeled with biotin-16-dUTP and visualized with HRP-anti-biotin. Most of TUNEL-positive follicle cells were found in methylation level of CCGG sites. Bar = 100 μm.

    Techniques Used: Methylation, TUNEL Assay, Staining, Labeling, Immunohistochemistry

    6) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    7) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    8) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    9) Product Images from "GLIS3, a Susceptibility Gene for Type 1 and Type 2 Diabetes, Modulates Pancreatic Beta Cell Apoptosis via Regulation of a Splice Variant of the BH3-Only Protein Bim"

    Article Title: GLIS3, a Susceptibility Gene for Type 1 and Type 2 Diabetes, Modulates Pancreatic Beta Cell Apoptosis via Regulation of a Splice Variant of the BH3-Only Protein Bim

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1003532

    GLIS3 regulates the differentiated beta cell phenotype. INS-1E cells, primary FACS-purified rat beta cells and human islet cells were transfected with control or GLIS3 siRNA (siCTL and siGLIS3, respectively; siGLIS3 here and below refers always to the GLIS3 siRNA#1; different siRNAs were used for rat and human cells as shown in Table S1 ). After 48 h, cells were used for real-time PCR analyses in INS-1E (A–E), primary rat beta cells (F–H), human islet cells (I) or functional studies (J–L). Results are means ± SEM corrected by the housekeeping genes GAPDH or β-actin (n = 4–5); (J) medium insulin accumulation of dispersed human islet cells; (K) glucose metabolism of INS-1E cells exposed to 1 or 10 mM glucose after GLIS3 KD (n = 6); (L) insulin secretion in INS-1E cells treated with 1 mM glucose, 10 mM glucose or 10 mM glucose plus forskolin (20 µM) after GLIS3 KD (n = 5). * P
    Figure Legend Snippet: GLIS3 regulates the differentiated beta cell phenotype. INS-1E cells, primary FACS-purified rat beta cells and human islet cells were transfected with control or GLIS3 siRNA (siCTL and siGLIS3, respectively; siGLIS3 here and below refers always to the GLIS3 siRNA#1; different siRNAs were used for rat and human cells as shown in Table S1 ). After 48 h, cells were used for real-time PCR analyses in INS-1E (A–E), primary rat beta cells (F–H), human islet cells (I) or functional studies (J–L). Results are means ± SEM corrected by the housekeeping genes GAPDH or β-actin (n = 4–5); (J) medium insulin accumulation of dispersed human islet cells; (K) glucose metabolism of INS-1E cells exposed to 1 or 10 mM glucose after GLIS3 KD (n = 6); (L) insulin secretion in INS-1E cells treated with 1 mM glucose, 10 mM glucose or 10 mM glucose plus forskolin (20 µM) after GLIS3 KD (n = 5). * P

    Techniques Used: FACS, Purification, Transfection, Real-time Polymerase Chain Reaction, Functional Assay

    GLIS3 KD potentiates cytokine-induced beta cell death via the mitochondrial pathway of apoptosis. INS-1E cells were transfected with siCTL or siGLIS3 (siGLIS3#1 and #2) and then exposed or not to cytokines. After 24 h cells were used for immunoblotting or immunofluorescence analysis. (A) Cleaved caspases-9 and -3. Blots are representative of 4 independent experiments. α-Tubulin was used as a control for protein loading in the different lanes; (B) cytochrome c release from the mitochondria to the cytosol. Blots are representative of 4 independent experiments. AIF and COX IV are used as mitochondrial markers, confirming adequate sub-cellular fractionation; (C) BAX localization was studied by immunocytochemistry. Nuclear morphology is shown by Hoechst staining. Arrows indicate BAX co-localization with ATP synthase β (mitochondrial marker). Images are representative of 4 independent experiments.
    Figure Legend Snippet: GLIS3 KD potentiates cytokine-induced beta cell death via the mitochondrial pathway of apoptosis. INS-1E cells were transfected with siCTL or siGLIS3 (siGLIS3#1 and #2) and then exposed or not to cytokines. After 24 h cells were used for immunoblotting or immunofluorescence analysis. (A) Cleaved caspases-9 and -3. Blots are representative of 4 independent experiments. α-Tubulin was used as a control for protein loading in the different lanes; (B) cytochrome c release from the mitochondria to the cytosol. Blots are representative of 4 independent experiments. AIF and COX IV are used as mitochondrial markers, confirming adequate sub-cellular fractionation; (C) BAX localization was studied by immunocytochemistry. Nuclear morphology is shown by Hoechst staining. Arrows indicate BAX co-localization with ATP synthase β (mitochondrial marker). Images are representative of 4 independent experiments.

    Techniques Used: Transfection, Immunofluorescence, Cell Fractionation, Immunocytochemistry, Staining, Marker

    Bim mediates the potentiation of apoptosis in GLIS3- deficient cells. INS-1E cells (A, D and E), primary rat beta cells (B) and human dispersed islet cells (C) transfected with control or GLIS3 siRNAs were exposed to cytokines (A, B, C and D) or palmitate (E) for 24 h. (A, B, C and E) cells transfected with Bim siRNA (siBim); (D) cells transfected with a second Bim siRNA affecting preferentially Bim S (siBim S ). Apoptosis was then measured using nuclear dyes. Results are means ± SEM (n = 4–11). * P
    Figure Legend Snippet: Bim mediates the potentiation of apoptosis in GLIS3- deficient cells. INS-1E cells (A, D and E), primary rat beta cells (B) and human dispersed islet cells (C) transfected with control or GLIS3 siRNAs were exposed to cytokines (A, B, C and D) or palmitate (E) for 24 h. (A, B, C and E) cells transfected with Bim siRNA (siBim); (D) cells transfected with a second Bim siRNA affecting preferentially Bim S (siBim S ). Apoptosis was then measured using nuclear dyes. Results are means ± SEM (n = 4–11). * P

    Techniques Used: Transfection

    10) Product Images from "Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes"

    Article Title: Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes

    Journal: bioRxiv

    doi: 10.1101/2020.02.26.966374

    HILPDA and DGAT1/DGAT2 colocalize and physically interact intracellularly. HepG2 cells were transfected with HILPDA_mEGFP and DGAT1_mCherry or DGAT2_mCherry under lipid loaded conditions. Microscopy was carried out on live cells. a) HILPDA_EGFP and mDGAT1_mCherry partially colocalize in HepG2 cells. b) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor DGAT1_mCherry. c) Intensity image and LUT coloured lifetime image from red (1300 ps) to blue (2800 ps) from HILPDA_EGFP lifetime (τ) in the absence (left) or presence (right) of DGAT1_mCherry indicating where interaction occurs d) HILPDA_EGFP and DGAT2_mCherry partially colocalize in HepG2 cells. e) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor DGAT2_mCherry. f) Intensity image and LUT coloured lifetime image from red (1300 ps) to blue (2800 ps) from HILPDA_EGFP lifetime (τ) in the absence (left) or presence (right) of DGAT2_mCherry indicating where interaction occurs g) HILPDA_EGFP and PLIN2_mCherry do not colocalize in HepG2 cells. h) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor PLIN2_mCherry. Asterisk indicates significantly different from donor only according to Student’s t test; **P
    Figure Legend Snippet: HILPDA and DGAT1/DGAT2 colocalize and physically interact intracellularly. HepG2 cells were transfected with HILPDA_mEGFP and DGAT1_mCherry or DGAT2_mCherry under lipid loaded conditions. Microscopy was carried out on live cells. a) HILPDA_EGFP and mDGAT1_mCherry partially colocalize in HepG2 cells. b) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor DGAT1_mCherry. c) Intensity image and LUT coloured lifetime image from red (1300 ps) to blue (2800 ps) from HILPDA_EGFP lifetime (τ) in the absence (left) or presence (right) of DGAT1_mCherry indicating where interaction occurs d) HILPDA_EGFP and DGAT2_mCherry partially colocalize in HepG2 cells. e) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor DGAT2_mCherry. f) Intensity image and LUT coloured lifetime image from red (1300 ps) to blue (2800 ps) from HILPDA_EGFP lifetime (τ) in the absence (left) or presence (right) of DGAT2_mCherry indicating where interaction occurs g) HILPDA_EGFP and PLIN2_mCherry do not colocalize in HepG2 cells. h) Fluorescence lifetime (τ) of HILPDA_EGFP in absence and presence of acceptor PLIN2_mCherry. Asterisk indicates significantly different from donor only according to Student’s t test; **P

    Techniques Used: Transfection, Microscopy, Fluorescence

    Hilpda expression is induced by fatty acids in various hepatoma cell lines. a) Venn diagram of upregulated genes (fold change > 1.5) in murine Hepa1-6 hepatoma cells treated with different fatty acids (500 µM) for 6h. b) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6 cells treated with different fatty acids (500 µM) for 6h. c) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6, Fao and HepG2 hepatoma cells treated for 24h with a 2:1 mixture of oleate and palmitate (total concentration 1.2 mM). d) Hilpda mRNA expression in wildtype mouse primary hepatocytes treated with oleate (500 µM) for 24h. Bar graphs are presented as mean ±SD. Asterisk indicates significantly different from control-treated cells according to Student’s t test; **P
    Figure Legend Snippet: Hilpda expression is induced by fatty acids in various hepatoma cell lines. a) Venn diagram of upregulated genes (fold change > 1.5) in murine Hepa1-6 hepatoma cells treated with different fatty acids (500 µM) for 6h. b) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6 cells treated with different fatty acids (500 µM) for 6h. c) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6, Fao and HepG2 hepatoma cells treated for 24h with a 2:1 mixture of oleate and palmitate (total concentration 1.2 mM). d) Hilpda mRNA expression in wildtype mouse primary hepatocytes treated with oleate (500 µM) for 24h. Bar graphs are presented as mean ±SD. Asterisk indicates significantly different from control-treated cells according to Student’s t test; **P

    Techniques Used: Expressing, Concentration Assay

    HILPDA promotes LD storage and increases DGAT1 levels in HepG2 cells. HepG2 cells were transduced with AV- Hilpda , AV- GFP , or non-transduced and treated with oleate:palmitate (2:1 ratio). a) HILPDA protein levels. b) Triglyceride content in cells incubated with serum free DMEM or 3h in 1 Mm oleate:palmitate. c) GFP fluorescence and BODIPY 493/503 staining. d) Quantification of LD size in HepG2 treated with 0.8mM oleate:palmitate for 8h and Hepa 1-6 cells treated with 1mM for 24h. e) LC3-I and LC3-II protein levels in HepG2 cells lipid loaded with 0.8mM oleate:palmitate for 8h, in the presence and absence of lysosomal inhibitors cocktail. f) Total DAG levels as determined by lipidomics in HepG2 cells incubated with 0.8mM oleate:palmitate for 5h. g) DGAT activity in HepG2 cells the presence and absence of the ATGL inhibitor Atglistatin. h) DGAT1 and HILPDA protein levels. i) mRNA levels of selected genes. j) DGAT1 protein levels in presence and absence of Atglistatin. j) DGAT1 and HILPDA protein levels in livers of mice infected with AAV- Gfp or AAV- Hilpda ( 21 ).
    Figure Legend Snippet: HILPDA promotes LD storage and increases DGAT1 levels in HepG2 cells. HepG2 cells were transduced with AV- Hilpda , AV- GFP , or non-transduced and treated with oleate:palmitate (2:1 ratio). a) HILPDA protein levels. b) Triglyceride content in cells incubated with serum free DMEM or 3h in 1 Mm oleate:palmitate. c) GFP fluorescence and BODIPY 493/503 staining. d) Quantification of LD size in HepG2 treated with 0.8mM oleate:palmitate for 8h and Hepa 1-6 cells treated with 1mM for 24h. e) LC3-I and LC3-II protein levels in HepG2 cells lipid loaded with 0.8mM oleate:palmitate for 8h, in the presence and absence of lysosomal inhibitors cocktail. f) Total DAG levels as determined by lipidomics in HepG2 cells incubated with 0.8mM oleate:palmitate for 5h. g) DGAT activity in HepG2 cells the presence and absence of the ATGL inhibitor Atglistatin. h) DGAT1 and HILPDA protein levels. i) mRNA levels of selected genes. j) DGAT1 protein levels in presence and absence of Atglistatin. j) DGAT1 and HILPDA protein levels in livers of mice infected with AAV- Gfp or AAV- Hilpda ( 21 ).

    Techniques Used: Transduction, Incubation, Fluorescence, Staining, Activity Assay, Mouse Assay, Infection

    HILPDA is primarily localized to the perinuclear area and preferentially associated with new fat. a) STED microscopy of HepG2 cells transfected with HILPDA fused to sYFP2 and lipid loaded with 0.8 mM oleate and 15 µM BODIPY C12 558/568 for 18h. λ ex : 470 nm (YFP) and 558 nm (BODIPY-558/568). λ em : 480-540 nm (sYFP2) and 570-650 nm (BODIPY 558/568). Left panel: HILPDA-sYFP2; middle panel: BODIPY-558, right panel: overlay. b) HepG2 cells cotransfected with HILPDA fused to Turquoise2 and with ER marker pDsRed2-ER (ClonTech) followed by incubation with 0.8mM oleate overnight.Left panel: HILPDA-Turquoise2; middle panel: ER marker pDsRed2-ER, right panel: overlay. c) Confocal microscopy of HepG2 cells transfected with HILPDA fused to Turquoise2 and lipid loaded with 0.6 mM oleate and 15 µM BODIPY C12 558/568 for 16h, followed by incubation for 20 min with BODIPY FL C12 and fixed with 3.7% PFA. λ ex : 440 nm (mTurquoise2), 561 nm (BODIPY 558/568), and 488 nm (BODIPY FL). λ em : 450-480 nm (mTurquoise2), 570-620 nm (BODIPY 558/568), and 505-558 nm (BODIPY FL). Colocalized pixels of HILPDA and Fluorescent fatty acids are represented on gray scale, higher colocalization is depicted with lighter pixels; non-colocalized HILPDA pixels are coloured green; whereas non-colocalized fluorescent fatty acid pixels are coloured red. d) Schematic depiction of the set-up and outcomes of the above experiments.
    Figure Legend Snippet: HILPDA is primarily localized to the perinuclear area and preferentially associated with new fat. a) STED microscopy of HepG2 cells transfected with HILPDA fused to sYFP2 and lipid loaded with 0.8 mM oleate and 15 µM BODIPY C12 558/568 for 18h. λ ex : 470 nm (YFP) and 558 nm (BODIPY-558/568). λ em : 480-540 nm (sYFP2) and 570-650 nm (BODIPY 558/568). Left panel: HILPDA-sYFP2; middle panel: BODIPY-558, right panel: overlay. b) HepG2 cells cotransfected with HILPDA fused to Turquoise2 and with ER marker pDsRed2-ER (ClonTech) followed by incubation with 0.8mM oleate overnight.Left panel: HILPDA-Turquoise2; middle panel: ER marker pDsRed2-ER, right panel: overlay. c) Confocal microscopy of HepG2 cells transfected with HILPDA fused to Turquoise2 and lipid loaded with 0.6 mM oleate and 15 µM BODIPY C12 558/568 for 16h, followed by incubation for 20 min with BODIPY FL C12 and fixed with 3.7% PFA. λ ex : 440 nm (mTurquoise2), 561 nm (BODIPY 558/568), and 488 nm (BODIPY FL). λ em : 450-480 nm (mTurquoise2), 570-620 nm (BODIPY 558/568), and 505-558 nm (BODIPY FL). Colocalized pixels of HILPDA and Fluorescent fatty acids are represented on gray scale, higher colocalization is depicted with lighter pixels; non-colocalized HILPDA pixels are coloured green; whereas non-colocalized fluorescent fatty acid pixels are coloured red. d) Schematic depiction of the set-up and outcomes of the above experiments.

    Techniques Used: Microscopy, Transfection, Marker, Incubation, Confocal Microscopy

    11) Product Images from "Silencing branched-chain ketoacid dehydrogenase or treatment with branched-chain ketoacids ex vivo inhibits muscle insulin signaling"

    Article Title: Silencing branched-chain ketoacid dehydrogenase or treatment with branched-chain ketoacids ex vivo inhibits muscle insulin signaling

    Journal: bioRxiv

    doi: 10.1101/2020.02.21.960153

    BCKAs impair insulin signaling in cardiomyocytes. a) Schematic of isolation of neonatal (NRCM) and adult (ARCM) rat cardiomyocytes and the treatment protocol. b) NRCMs and c) ARCMs were pre-incubated with either 2% BSA or 2% BSA conjugated with 0.4mM palmitate for 16 hr followed by 5mM ketoleucine for 30 min. Immunoblot and densitometric analysis of total and phosphorylated AKT Ser 473 levels in cardiomyocytes subjected to either 200 nM (NRCMs) or 100 nM (ARCMs) insulin stimulation for 20 mins in the presence or absence of ketoleucine. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p
    Figure Legend Snippet: BCKAs impair insulin signaling in cardiomyocytes. a) Schematic of isolation of neonatal (NRCM) and adult (ARCM) rat cardiomyocytes and the treatment protocol. b) NRCMs and c) ARCMs were pre-incubated with either 2% BSA or 2% BSA conjugated with 0.4mM palmitate for 16 hr followed by 5mM ketoleucine for 30 min. Immunoblot and densitometric analysis of total and phosphorylated AKT Ser 473 levels in cardiomyocytes subjected to either 200 nM (NRCMs) or 100 nM (ARCMs) insulin stimulation for 20 mins in the presence or absence of ketoleucine. Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test; *p

    Techniques Used: Isolation, Incubation

    12) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    13) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    14) Product Images from "Basal hypersecretion of glucagon and insulin from palmitate-exposed human islets depends on FFAR1 but not decreased somatostatin secretion"

    Article Title: Basal hypersecretion of glucagon and insulin from palmitate-exposed human islets depends on FFAR1 but not decreased somatostatin secretion

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-04730-5

    Role of FFAR1 and fatty acid β-oxidation in the effect of palmitate on EndoC-βH1 insulin secretion and respiration. Cells were exposed to 5.6 mM glucose in the absence (white symbols) or presence (black symbols) of palmitate with and without ANT825 and/or etomoxir, trimetazidine. Insulin secretion (panel A). Representative kinetic OCR recordings from cells acutely exposed to palmitate with and without ANT825 and/or etomoxir (panel B). Mitochondrial OCR (panel C), ATP-coupled OCR (panel D) and proton leak OCR (panel E). Results show mean ± SEM (panels A,C-E,) (n = 5), and mean ± SD (panel B) (N = 6). *P
    Figure Legend Snippet: Role of FFAR1 and fatty acid β-oxidation in the effect of palmitate on EndoC-βH1 insulin secretion and respiration. Cells were exposed to 5.6 mM glucose in the absence (white symbols) or presence (black symbols) of palmitate with and without ANT825 and/or etomoxir, trimetazidine. Insulin secretion (panel A). Representative kinetic OCR recordings from cells acutely exposed to palmitate with and without ANT825 and/or etomoxir (panel B). Mitochondrial OCR (panel C), ATP-coupled OCR (panel D) and proton leak OCR (panel E). Results show mean ± SEM (panels A,C-E,) (n = 5), and mean ± SD (panel B) (N = 6). *P

    Techniques Used:

    15) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    16) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    17) Product Images from "Lysophosphatidic Acid Up-Regulates Hexokinase II and Glycolysis to Promote Proliferation of Ovarian Cancer Cells"

    Article Title: Lysophosphatidic Acid Up-Regulates Hexokinase II and Glycolysis to Promote Proliferation of Ovarian Cancer Cells

    Journal: Neoplasia (New York, N.Y.)

    doi: 10.1016/j.neo.2015.09.003

    LPA stimulates expression of HK2. (A) Caov-3 cells were treated with LPA (10 μM) or vehicle (BSA) for 12 hours. The human glucose metabolism RT 2 profiler PCR array was performed on cDNAs prepared from total cellular RNA as described in Experimental Procedures. Expression levels of glycolytic genes were normalized on GAPDH and then compared between LPA-treated and control cells and presented as fold changes over vehicle controls (defined as arbitrary 1). (B, C) Caov-3 and OVCA-432 cells were treated with LPA (10 μM) for the indicated periods of time (hours). Induction of HK2 mRNA (B) and protein (C) was analyzed by RT-qPCR and immunoblotting, respectively. HK2 protein levels were quantified by densitometry and presented as fold changes relative to control cells. (D) Hexokinase activity was determined in Caov-3 and OVCA-432 cells treated for 16 hours with LPA or vehicle and presented as nanomoles/mg protein/min at 37 °C. (E) HK-2 expression was down-regulated by HK2 specific siRNA in Caov-3 and OVCA-432 cells. LPA-induced glycolysis in HK-2 knockdown and control siRNA-treated cells were determined as described in Figure 1 .
    Figure Legend Snippet: LPA stimulates expression of HK2. (A) Caov-3 cells were treated with LPA (10 μM) or vehicle (BSA) for 12 hours. The human glucose metabolism RT 2 profiler PCR array was performed on cDNAs prepared from total cellular RNA as described in Experimental Procedures. Expression levels of glycolytic genes were normalized on GAPDH and then compared between LPA-treated and control cells and presented as fold changes over vehicle controls (defined as arbitrary 1). (B, C) Caov-3 and OVCA-432 cells were treated with LPA (10 μM) for the indicated periods of time (hours). Induction of HK2 mRNA (B) and protein (C) was analyzed by RT-qPCR and immunoblotting, respectively. HK2 protein levels were quantified by densitometry and presented as fold changes relative to control cells. (D) Hexokinase activity was determined in Caov-3 and OVCA-432 cells treated for 16 hours with LPA or vehicle and presented as nanomoles/mg protein/min at 37 °C. (E) HK-2 expression was down-regulated by HK2 specific siRNA in Caov-3 and OVCA-432 cells. LPA-induced glycolysis in HK-2 knockdown and control siRNA-treated cells were determined as described in Figure 1 .

    Techniques Used: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Activity Assay

    18) Product Images from "Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes"

    Article Title: Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes

    Journal: bioRxiv

    doi: 10.1101/2020.02.26.966374

    HILPDA promotes LD storage and increases DGAT1 levels in HepG2 cells. HepG2 cells were transduced with AV- Hilpda , AV- GFP , or non-transduced and treated with oleate:palmitate (2:1 ratio). a) HILPDA protein levels. b) Triglyceride content in cells incubated with serum free DMEM or 3h in 1 Mm oleate:palmitate. c) GFP fluorescence and BODIPY 493/503 staining. d) Quantification of LD size in HepG2 treated with 0.8mM oleate:palmitate for 8h and Hepa 1-6 cells treated with 1mM for 24h. e) LC3-I and LC3-II protein levels in HepG2 cells lipid loaded with 0.8mM oleate:palmitate for 8h, in the presence and absence of lysosomal inhibitors cocktail. f) Total DAG levels as determined by lipidomics in HepG2 cells incubated with 0.8mM oleate:palmitate for 5h. g) DGAT activity in HepG2 cells the presence and absence of the ATGL inhibitor Atglistatin. h) DGAT1 and HILPDA protein levels. i) mRNA levels of selected genes. j) DGAT1 protein levels in presence and absence of Atglistatin. j) DGAT1 and HILPDA protein levels in livers of mice infected with AAV- Gfp or AAV- Hilpda ( 21 ).
    Figure Legend Snippet: HILPDA promotes LD storage and increases DGAT1 levels in HepG2 cells. HepG2 cells were transduced with AV- Hilpda , AV- GFP , or non-transduced and treated with oleate:palmitate (2:1 ratio). a) HILPDA protein levels. b) Triglyceride content in cells incubated with serum free DMEM or 3h in 1 Mm oleate:palmitate. c) GFP fluorescence and BODIPY 493/503 staining. d) Quantification of LD size in HepG2 treated with 0.8mM oleate:palmitate for 8h and Hepa 1-6 cells treated with 1mM for 24h. e) LC3-I and LC3-II protein levels in HepG2 cells lipid loaded with 0.8mM oleate:palmitate for 8h, in the presence and absence of lysosomal inhibitors cocktail. f) Total DAG levels as determined by lipidomics in HepG2 cells incubated with 0.8mM oleate:palmitate for 5h. g) DGAT activity in HepG2 cells the presence and absence of the ATGL inhibitor Atglistatin. h) DGAT1 and HILPDA protein levels. i) mRNA levels of selected genes. j) DGAT1 protein levels in presence and absence of Atglistatin. j) DGAT1 and HILPDA protein levels in livers of mice infected with AAV- Gfp or AAV- Hilpda ( 21 ).

    Techniques Used: Transduction, Incubation, Fluorescence, Staining, Activity Assay, Mouse Assay, Infection

    19) Product Images from "TPD52 expression increases neutral lipid storage within cultured cells"

    Article Title: TPD52 expression increases neutral lipid storage within cultured cells

    Journal: Journal of Cell Science

    doi: 10.1242/jcs.167692

    Colocalisation of TPD52 and the Golgi marker Gm130 in untreated and oleic-acid-treated D52-2-7 cells. Cells were untreated (A) or supplemented with 400 μM oleic acid in fatty-acid-free BSA (OA) for 6 h (B), fixed and subjected
    Figure Legend Snippet: Colocalisation of TPD52 and the Golgi marker Gm130 in untreated and oleic-acid-treated D52-2-7 cells. Cells were untreated (A) or supplemented with 400 μM oleic acid in fatty-acid-free BSA (OA) for 6 h (B), fixed and subjected

    Techniques Used: Marker

    Colocalisation of Gm130 or TPD52 with ARL1 and ADRP in untreated and oleic-acid-treated D52-2-7 cells. Cells were untreated or supplemented with 400 μM in fatty-acid-free BSA (OA) for 24 h, fixed, and subjected to indirect immunofluorescence
    Figure Legend Snippet: Colocalisation of Gm130 or TPD52 with ARL1 and ADRP in untreated and oleic-acid-treated D52-2-7 cells. Cells were untreated or supplemented with 400 μM in fatty-acid-free BSA (OA) for 24 h, fixed, and subjected to indirect immunofluorescence

    Techniques Used: Immunofluorescence

    20) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    21) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    22) Product Images from "Effects of LPS and dietary free fatty acids on MCP-1 in 3T3-L1 adipocytes and macrophages in vitro"

    Article Title: Effects of LPS and dietary free fatty acids on MCP-1 in 3T3-L1 adipocytes and macrophages in vitro

    Journal: Nutrition & Diabetes

    doi: 10.1038/nutd.2014.10

    Effects of ( a ) FFA and LPS stimulation on MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and 500 μ M of BSA-complexed fatty acid for 24 h. The fatty acids investigated are: monounsaturated fatty acid (oleic acid/OA), saturated fatty acid (palmitic acid/PA) and trans fatty acid (elaidic acid/EA). Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), ns=non-significant vs FAF-BSA.
    Figure Legend Snippet: Effects of ( a ) FFA and LPS stimulation on MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and 500 μ M of BSA-complexed fatty acid for 24 h. The fatty acids investigated are: monounsaturated fatty acid (oleic acid/OA), saturated fatty acid (palmitic acid/PA) and trans fatty acid (elaidic acid/EA). Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), ns=non-significant vs FAF-BSA.

    Techniques Used: Expressing, Incubation

    Effect of palmitic acid on MCP-1 mRNA expression in THP-1 macrophages. THP-1 macrophages were incubated with RPMI-1640 growth media containing 2% FAF-BSA and palmitic acid (PA, 500 μ M ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Effect of palmitic acid on MCP-1 mRNA expression in THP-1 macrophages. THP-1 macrophages were incubated with RPMI-1640 growth media containing 2% FAF-BSA and palmitic acid (PA, 500 μ M ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    Dose-dependent effects of LPS on MCP-1 mRNA expression in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 to 500 ng ml −1 ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Dose-dependent effects of LPS on MCP-1 mRNA expression in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 to 500 ng ml −1 ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    Effect of LPS and a TLR4-inhibitor on ( a ) TLR4 and MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Effect of LPS and a TLR4-inhibitor on ( a ) TLR4 and MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    23) Product Images from "Supported Bilayers with Excess Membrane Reservoir: A Template for Reconstituting Membrane Budding and Fission"

    Article Title: Supported Bilayers with Excess Membrane Reservoir: A Template for Reconstituting Membrane Budding and Fission

    Journal: Biophysical Journal

    doi: 10.1016/j.bpj.2010.04.036

    ( A ) Triton X-100-induced solubilization of the membrane bilayer on templates. Templates were formed with DOPC/DOPS/RhPE (84:15:1 mol%) SUVs in the presence of 1 M NaCl. RhPE fluorescence released into solution with increasing concentrations of Triton
    Figure Legend Snippet: ( A ) Triton X-100-induced solubilization of the membrane bilayer on templates. Templates were formed with DOPC/DOPS/RhPE (84:15:1 mol%) SUVs in the presence of 1 M NaCl. RhPE fluorescence released into solution with increasing concentrations of Triton

    Techniques Used: Fluorescence

    24) Product Images from "Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary"

    Article Title: Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.17039

    Methylation level of CCGG sites in TUNEL-positive follicle cells in adjacent sections of mouse ovary. Adjacent sections of paraffin-embedded mouse ovary were used for H E staining, TUNEL, and the detection of non-methylated CCGG sites and methylated CCGG sites, as described in detail in “II. Materials and Methods”. A : H E staining. B : TUNEL staining. C : Blockade of 3'-OH ends with dideoxynucleotides by TdT. After blockade, the section was labeled with biotin-16-dUTP by TdT and the incorporated biotin was detected with HRP-anti-biotin. No signals were observed. D : Staining for non-methylated CCGG sites. After the blockade procedure described in ( C ), the section was digested with Hpa II, labeled with biotin-16-dUTP and visualized by enzyme-immunohistochemistry with HRP-anti-biotin. E : Blockade of Hpa II cutting sites with dideoxynucleotides by TdT. The section was digested with Hpa II and the cutting sites were blocked with a dideoxynucleotide mixture. Then the section was processed in a manner similar to that described in ( C ). F : Staining for methylated CCGG sites. After blockade of Hpa II cutting sites with dideoxynucleotides, the section was digested with Msp I and the cutting sites were labeled with biotin-16-dUTP and visualized with HRP-anti-biotin. Most of TUNEL-positive follicle cells were found in methylation level of CCGG sites. Bar = 100 μm.
    Figure Legend Snippet: Methylation level of CCGG sites in TUNEL-positive follicle cells in adjacent sections of mouse ovary. Adjacent sections of paraffin-embedded mouse ovary were used for H E staining, TUNEL, and the detection of non-methylated CCGG sites and methylated CCGG sites, as described in detail in “II. Materials and Methods”. A : H E staining. B : TUNEL staining. C : Blockade of 3'-OH ends with dideoxynucleotides by TdT. After blockade, the section was labeled with biotin-16-dUTP by TdT and the incorporated biotin was detected with HRP-anti-biotin. No signals were observed. D : Staining for non-methylated CCGG sites. After the blockade procedure described in ( C ), the section was digested with Hpa II, labeled with biotin-16-dUTP and visualized by enzyme-immunohistochemistry with HRP-anti-biotin. E : Blockade of Hpa II cutting sites with dideoxynucleotides by TdT. The section was digested with Hpa II and the cutting sites were blocked with a dideoxynucleotide mixture. Then the section was processed in a manner similar to that described in ( C ). F : Staining for methylated CCGG sites. After blockade of Hpa II cutting sites with dideoxynucleotides, the section was digested with Msp I and the cutting sites were labeled with biotin-16-dUTP and visualized with HRP-anti-biotin. Most of TUNEL-positive follicle cells were found in methylation level of CCGG sites. Bar = 100 μm.

    Techniques Used: Methylation, TUNEL Assay, Staining, Labeling, Immunohistochemistry

    Simultaneous localization of non-methylated and methylated GATCG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Sau 3A I cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Mbo I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images are given in C, F and I . Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated GATCG was localized in the nuclei of granulosa cells (white arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.
    Figure Legend Snippet: Simultaneous localization of non-methylated and methylated GATCG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Sau 3A I cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Mbo I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images are given in C, F and I . Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated GATCG was localized in the nuclei of granulosa cells (white arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.

    Techniques Used: Methylation, Labeling, Staining

    Simultaneous localization of non-methylated and methylated CCGG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Hpa II cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Msp I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images were shown in C, F and I . A, B and C : Primary follicle . D, E and F : Secondary follicle. G, H and I : Tertiary follicle. Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated CCGG was localized in the nuclei of granulosa cells (white arrows) or oocytes (red arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.
    Figure Legend Snippet: Simultaneous localization of non-methylated and methylated CCGG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Hpa II cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Msp I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin ( A, D and G ) and rhodamine-anti-digoxigenin ( B, E and H ), respectively. Merged images were shown in C, F and I . A, B and C : Primary follicle . D, E and F : Secondary follicle. G, H and I : Tertiary follicle. Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated CCGG was localized in the nuclei of granulosa cells (white arrows) or oocytes (red arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.

    Techniques Used: Methylation, Labeling, Staining

    25) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    26) Product Images from "Effects of LPS and dietary free fatty acids on MCP-1 in 3T3-L1 adipocytes and macrophages in vitro"

    Article Title: Effects of LPS and dietary free fatty acids on MCP-1 in 3T3-L1 adipocytes and macrophages in vitro

    Journal: Nutrition & Diabetes

    doi: 10.1038/nutd.2014.10

    Effects of ( a ) FFA and LPS stimulation on MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and 500 μ M of BSA-complexed fatty acid for 24 h. The fatty acids investigated are: monounsaturated fatty acid (oleic acid/OA), saturated fatty acid (palmitic acid/PA) and trans fatty acid (elaidic acid/EA). Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), ns=non-significant vs FAF-BSA.
    Figure Legend Snippet: Effects of ( a ) FFA and LPS stimulation on MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and 500 μ M of BSA-complexed fatty acid for 24 h. The fatty acids investigated are: monounsaturated fatty acid (oleic acid/OA), saturated fatty acid (palmitic acid/PA) and trans fatty acid (elaidic acid/EA). Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), ns=non-significant vs FAF-BSA.

    Techniques Used: Expressing, Incubation

    Effect of palmitic acid on MCP-1 mRNA expression in THP-1 macrophages. THP-1 macrophages were incubated with RPMI-1640 growth media containing 2% FAF-BSA and palmitic acid (PA, 500 μ M ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Effect of palmitic acid on MCP-1 mRNA expression in THP-1 macrophages. THP-1 macrophages were incubated with RPMI-1640 growth media containing 2% FAF-BSA and palmitic acid (PA, 500 μ M ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    Dose-dependent effects of LPS on MCP-1 mRNA expression in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 to 500 ng ml −1 ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Dose-dependent effects of LPS on MCP-1 mRNA expression in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 to 500 ng ml −1 ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    Effect of LPS and a TLR4-inhibitor on ( a ) TLR4 and MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P
    Figure Legend Snippet: Effect of LPS and a TLR4-inhibitor on ( a ) TLR4 and MCP-1 mRNA expression and ( b ) MCP-1 protein secretion in 3T3-L1 adipocytes. 3T3-L1 adipocytes were incubated with DMEM growth media containing 2% FAF-BSA and LPS (2 ng ml −1 ) alone or in combination with the TLR4-inhibitor CLI095 (CLI, 3 μ M ) for 24 h. Results were expressed as ±s.e.m., relative to FAF-BSA=1 ( n =9), * P

    Techniques Used: Expressing, Incubation

    27) Product Images from "Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes"

    Article Title: Hypoxia-inducible lipid droplet-associated interacts with DGAT1 to promote lipid storage in hepatocytes

    Journal: bioRxiv

    doi: 10.1101/2020.02.26.966374

    Hilpda expression is induced by fatty acids in various hepatoma cell lines. a) Venn diagram of upregulated genes (fold change > 1.5) in murine Hepa1-6 hepatoma cells treated with different fatty acids (500 µM) for 6h. b) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6 cells treated with different fatty acids (500 µM) for 6h. c) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6, Fao and HepG2 hepatoma cells treated for 24h with a 2:1 mixture of oleate and palmitate (total concentration 1.2 mM). d) Hilpda mRNA expression in wildtype mouse primary hepatocytes treated with oleate (500 µM) for 24h. Bar graphs are presented as mean ±SD. Asterisk indicates significantly different from control-treated cells according to Student’s t test; **P
    Figure Legend Snippet: Hilpda expression is induced by fatty acids in various hepatoma cell lines. a) Venn diagram of upregulated genes (fold change > 1.5) in murine Hepa1-6 hepatoma cells treated with different fatty acids (500 µM) for 6h. b) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6 cells treated with different fatty acids (500 µM) for 6h. c) Relative changes in Hilpda and Plin2 mRNA in Hepa1-6, Fao and HepG2 hepatoma cells treated for 24h with a 2:1 mixture of oleate and palmitate (total concentration 1.2 mM). d) Hilpda mRNA expression in wildtype mouse primary hepatocytes treated with oleate (500 µM) for 24h. Bar graphs are presented as mean ±SD. Asterisk indicates significantly different from control-treated cells according to Student’s t test; **P

    Techniques Used: Expressing, Concentration Assay

    28) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    29) Product Images from "Lysophosphatidic Acid Activates Lipogenic Pathways and de Novo Lipid Synthesis in Ovarian Cancer Cells *"

    Article Title: Lysophosphatidic Acid Activates Lipogenic Pathways and de Novo Lipid Synthesis in Ovarian Cancer Cells *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M112.340083

    LPA stimulates de novo lipid synthesis. Caov-3, OVCA-432, and IOSE-29 cells were treated with LPA (10 μ m ) or BSA (vehicle) for 24 h. In the last 6 h of incubation, the cells were pulse-labeled with 5 μCi/ml of [ 14 C]acetic acid before lipid
    Figure Legend Snippet: LPA stimulates de novo lipid synthesis. Caov-3, OVCA-432, and IOSE-29 cells were treated with LPA (10 μ m ) or BSA (vehicle) for 24 h. In the last 6 h of incubation, the cells were pulse-labeled with 5 μCi/ml of [ 14 C]acetic acid before lipid

    Techniques Used: Incubation, Labeling

    30) Product Images from "Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria"

    Article Title: Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria

    Journal: Scientific Reports

    doi: 10.1038/srep37798

    Ca 2+ retention capacity of freeze-thawed Hela S3 human mitochondria. Hela S3 mitochondria (0.5 mg protein ml −1 ) were incubated with Fluo-4FF (0.35 μM) in the presence of succinate (10 mM) and rotenone (1 μM). Pulses of 5 μM CaCl 2 were sequentially added at 3 minute intervals and extra-mitochondrial Fluo-4FF fluorescence measured. Representative raw traces of Ca 2+ uptake in the presence of ( a ) ER-000444793 and ( b ) CsA demonstrate complete Ca 2+ uptake prior to concentration-dependent Ca 2+ -induced mPT. ( c,d ) Area under the curve was calculated between 60–660 seconds and results are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). Data are expressed as means (± s.d.) of at least three independent experiments. Curve fitting used a 4-parameter logistic equation (GraphPad Prism). Raw traces are representative of at least three independent experiments. Abbreviations: CsA; cyclosporin A, a.u; arbitrary unit.
    Figure Legend Snippet: Ca 2+ retention capacity of freeze-thawed Hela S3 human mitochondria. Hela S3 mitochondria (0.5 mg protein ml −1 ) were incubated with Fluo-4FF (0.35 μM) in the presence of succinate (10 mM) and rotenone (1 μM). Pulses of 5 μM CaCl 2 were sequentially added at 3 minute intervals and extra-mitochondrial Fluo-4FF fluorescence measured. Representative raw traces of Ca 2+ uptake in the presence of ( a ) ER-000444793 and ( b ) CsA demonstrate complete Ca 2+ uptake prior to concentration-dependent Ca 2+ -induced mPT. ( c,d ) Area under the curve was calculated between 60–660 seconds and results are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). Data are expressed as means (± s.d.) of at least three independent experiments. Curve fitting used a 4-parameter logistic equation (GraphPad Prism). Raw traces are representative of at least three independent experiments. Abbreviations: CsA; cyclosporin A, a.u; arbitrary unit.

    Techniques Used: Incubation, Fluorescence, Concentration Assay, Inhibition

    Response of freeze-thawed mitochondria to Ca 2+ -induced mPT. Rat liver mitochondria (2 mg protein ml −1 ) were incubated with TMRM (5 μM) in the presence of succinate (10 mM) and rotenone (1 μM). ( a ) TMRM fluorescence pre-read (ex. 560/em. 590 nm) revealed no significant effect on ER-000444793 (50 μM) or CsA (10 μM) after 10 minutes incubation. Statistical significance was calculated using a one-way ANOVA corrected for multiple comparisons using Tukey method (NS P > 0.05; GraphPad Prism). Mitochondria were incubated with ( b ) ER-000444793 or ( c ) CsA in a 2-fold dilution series for 10 minutes prior to the addition of a CaCl 2 bolus (150 μM). TMRM fluorescence was measured after 20 minutes incubation. Results are expressed as % depolarisation, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). ( d–g ) Rat liver mitochondria (1 mg protein ml −1 ) were incubated with Fluo-4FF (0.35 μM) in the presence of succinate (10 mM) and rotenone (1 μM). Pulses of CaCl 2 (10 μM) were added sequentially and extra-mitochondrial fluorescence measured. Representative raw traces of Ca 2+ uptake in the presence of ( d ) ER-000444793 and ( e ) CsA demonstrate complete Ca 2+ uptake prior to concentration-dependent, Ca 2+ -induced mPT. ( f,g ) Area under the curve was calculated between 240–840 seconds and results are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). Data are expressed as means (±s.d.) of at least three independent experiments. Curves fitting used a 4-parameter logistic equation (GraphPad Prism). Raw traces are representative of at least three independent experiments. Abbreviations: NS; not significant, CsA; cyclosporin A, TMRM; tetramethylrhodamine methylester. a.u; arbitrary unit.
    Figure Legend Snippet: Response of freeze-thawed mitochondria to Ca 2+ -induced mPT. Rat liver mitochondria (2 mg protein ml −1 ) were incubated with TMRM (5 μM) in the presence of succinate (10 mM) and rotenone (1 μM). ( a ) TMRM fluorescence pre-read (ex. 560/em. 590 nm) revealed no significant effect on ER-000444793 (50 μM) or CsA (10 μM) after 10 minutes incubation. Statistical significance was calculated using a one-way ANOVA corrected for multiple comparisons using Tukey method (NS P > 0.05; GraphPad Prism). Mitochondria were incubated with ( b ) ER-000444793 or ( c ) CsA in a 2-fold dilution series for 10 minutes prior to the addition of a CaCl 2 bolus (150 μM). TMRM fluorescence was measured after 20 minutes incubation. Results are expressed as % depolarisation, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). ( d–g ) Rat liver mitochondria (1 mg protein ml −1 ) were incubated with Fluo-4FF (0.35 μM) in the presence of succinate (10 mM) and rotenone (1 μM). Pulses of CaCl 2 (10 μM) were added sequentially and extra-mitochondrial fluorescence measured. Representative raw traces of Ca 2+ uptake in the presence of ( d ) ER-000444793 and ( e ) CsA demonstrate complete Ca 2+ uptake prior to concentration-dependent, Ca 2+ -induced mPT. ( f,g ) Area under the curve was calculated between 240–840 seconds and results are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). Data are expressed as means (±s.d.) of at least three independent experiments. Curves fitting used a 4-parameter logistic equation (GraphPad Prism). Raw traces are representative of at least three independent experiments. Abbreviations: NS; not significant, CsA; cyclosporin A, TMRM; tetramethylrhodamine methylester. a.u; arbitrary unit.

    Techniques Used: Incubation, Fluorescence, Inhibition, Concentration Assay

    31) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    32) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    33) Product Images from "Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA"

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    Journal:

    doi: 10.1016/j.metabol.2007.09.015

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Techniques Used:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Techniques Used: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol
    Figure Legend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Techniques Used: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets
    Figure Legend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Techniques Used: Isolation

    Related Articles

    Centrifugation:

    Article Title: Molecular Characterization of the Host Defense Activity of the Barrier to Autointegration Factor against Vaccinia Virus ▿
    Article Snippet: .. Lysis was performed for 10 min on ice in lysis buffer (50 mM Tris [pH 7.4], 75 mM NaCl, 1 mM EDTA, 4% sucrose, and 0.5% Triton X-100) supplemented with fresh 1 mM phenylmethylsulfonyl fluoride (PMSF) and protease and phosphatase inhibitors (Complete Mini and PhosStop tablets; Roche), after which nuclei were removed following centrifugation at 800 × g . .. For DNA binding assays, 50 μl of native dsDNA cellulose beads (Amersham) was added to the lysates and incubated overnight at 4°C with end-over-end rotation.

    Amplification:

    Article Title: Addition of a Single gp120 Glycan Confers Increased Binding to Dendritic Cell-Specific ICAM-3-Grabbing Nonintegrin and Neutralization Escape to Human Immunodeficiency Virus Type 1
    Article Snippet: .. Specific cDNAs were amplified according to the manufacturer's specifications in a 100-μl reaction mixture containing 5 μl of cDNA; 10 mM Tris-HCl (pH 8.5); 50 mM KCl; 1.5 mM MgCl2 ; 0.1% Triton X-100; 200 μM (each) dATP, dGTP, dCTP, and dTTP; 20 pmol of each primer; and 2.5 U of EXPAND high-fidelity DNA polymerase (Roche Diagnostic Corp., Indianapolis, Ind.). .. PCR primers hybridizing to the 5′- and 3′-untranslated regions of rhesus and human DC-SIGN were as follows (restriction sites for cloning are underlined): first round, DC-SIGNf1 (5′-TCT GGA CAC TGG GGG AGA GTG G-3′) and DC-SIGNb1 (5′-GGA TGG AGA GAA GGA ACT GTA G-3′); second round, DC-SIGNf2 (5′-TCGAG GGATCCGAATTC GGA GAG TGG GGT GAC ATG AGT G-3′) and DC-SIGNb2 (5′-TCGA GCGGCCGCTCTAGA GCT TAA AAG GGG GTG AAG TTC TG-3′).

    Protease Inhibitor:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Article Title: Identification of Domains and Residues within the ? Subunit of Eukaryotic Translation Initiation Factor 2B (eIF2B?) Required for Guanine Nucleotide Exchange Reveals a Novel Activation Function Promoted by eIF2B Complex Formation
    Article Snippet: .. Purified eIF2Bɛ proteins (200 nM), 100 nM purified eIF2B complex, or an equivalent concentration of control FLAG peptide was incubated with 20 μl (wet volume) of anti-FLAG M2 affinity resin (Eastman Kodak) with rotation for 2 h at 4°C in 100 μl of buffer A (100 mM KCl, 20 mM Tris-HCl [pH 7.5], 2 mM MgCl2 , 5 mM β-mercaptoethanol, 0.1% Triton X-100) in the presence of Complete EDTA-free protease inhibitor (Roche Diagnostics) and 10 μg of BSA. .. Beads were washed three times with 100 μl of buffer A. Purified eIF2 at concentrations of 0.625 to 40 nM (as indicated in the figure legends) was then added to the beads in 100 μl of buffer A in the presence of 10 μg of BSA and rotated for 2 h at 4°C.

    Article Title: Luteolin attenuates sepsis-induced myocardial injury by enhancing autophagy in mice
    Article Snippet: .. Western blot analysis Cardiac tissues or cardiomyocytes were harvested and homogenized with RIPA buffer containing protease inhibitor cocktail (Roche Diagnostics) on ice. .. After centrifugation at 12,000 × g for 15 min at 4°C, the total protein content of the supernatant was quantified using a bicinchoninic acid protein assay (Applygen Technologies, Inc.) and the supernatant was stored at −80°C until use.

    Transfection:

    Article Title: The Bovine Immunodeficiency Virus Rev Protein: Identification of a Novel Nuclear Import Pathway and Nuclear Export Signal among Retroviral Rev/Rev-Like Proteins
    Article Snippet: .. After an incubation of 48 h, the transfected cells were fixed with 4% paraformaldehyde in PBS for 15 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with 4% BSA in PBS for 1 h at 37°C, and then incubated with mouse primary anti-Myc antibodies (1 μg/ml) (Roche) for 1 h at 37°C. .. After three washes with PBS containing 0.2% Triton X-100, the cells were incubated with Alexa Fluor 488 -labeled anti-mouse secondary antibodies (Invitrogen) for 1 h at 37°C.

    Purification:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Article Title: Identification of Domains and Residues within the ? Subunit of Eukaryotic Translation Initiation Factor 2B (eIF2B?) Required for Guanine Nucleotide Exchange Reveals a Novel Activation Function Promoted by eIF2B Complex Formation
    Article Snippet: .. Purified eIF2Bɛ proteins (200 nM), 100 nM purified eIF2B complex, or an equivalent concentration of control FLAG peptide was incubated with 20 μl (wet volume) of anti-FLAG M2 affinity resin (Eastman Kodak) with rotation for 2 h at 4°C in 100 μl of buffer A (100 mM KCl, 20 mM Tris-HCl [pH 7.5], 2 mM MgCl2 , 5 mM β-mercaptoethanol, 0.1% Triton X-100) in the presence of Complete EDTA-free protease inhibitor (Roche Diagnostics) and 10 μg of BSA. .. Beads were washed three times with 100 μl of buffer A. Purified eIF2 at concentrations of 0.625 to 40 nM (as indicated in the figure legends) was then added to the beads in 100 μl of buffer A in the presence of 10 μg of BSA and rotated for 2 h at 4°C.

    Concentration Assay:

    Article Title: Identification of Domains and Residues within the ? Subunit of Eukaryotic Translation Initiation Factor 2B (eIF2B?) Required for Guanine Nucleotide Exchange Reveals a Novel Activation Function Promoted by eIF2B Complex Formation
    Article Snippet: .. Purified eIF2Bɛ proteins (200 nM), 100 nM purified eIF2B complex, or an equivalent concentration of control FLAG peptide was incubated with 20 μl (wet volume) of anti-FLAG M2 affinity resin (Eastman Kodak) with rotation for 2 h at 4°C in 100 μl of buffer A (100 mM KCl, 20 mM Tris-HCl [pH 7.5], 2 mM MgCl2 , 5 mM β-mercaptoethanol, 0.1% Triton X-100) in the presence of Complete EDTA-free protease inhibitor (Roche Diagnostics) and 10 μg of BSA. .. Beads were washed three times with 100 μl of buffer A. Purified eIF2 at concentrations of 0.625 to 40 nM (as indicated in the figure legends) was then added to the beads in 100 μl of buffer A in the presence of 10 μg of BSA and rotated for 2 h at 4°C.

    Incubation:

    Article Title: Identification of Domains and Residues within the ? Subunit of Eukaryotic Translation Initiation Factor 2B (eIF2B?) Required for Guanine Nucleotide Exchange Reveals a Novel Activation Function Promoted by eIF2B Complex Formation
    Article Snippet: .. Purified eIF2Bɛ proteins (200 nM), 100 nM purified eIF2B complex, or an equivalent concentration of control FLAG peptide was incubated with 20 μl (wet volume) of anti-FLAG M2 affinity resin (Eastman Kodak) with rotation for 2 h at 4°C in 100 μl of buffer A (100 mM KCl, 20 mM Tris-HCl [pH 7.5], 2 mM MgCl2 , 5 mM β-mercaptoethanol, 0.1% Triton X-100) in the presence of Complete EDTA-free protease inhibitor (Roche Diagnostics) and 10 μg of BSA. .. Beads were washed three times with 100 μl of buffer A. Purified eIF2 at concentrations of 0.625 to 40 nM (as indicated in the figure legends) was then added to the beads in 100 μl of buffer A in the presence of 10 μg of BSA and rotated for 2 h at 4°C.

    Article Title: The Bovine Immunodeficiency Virus Rev Protein: Identification of a Novel Nuclear Import Pathway and Nuclear Export Signal among Retroviral Rev/Rev-Like Proteins
    Article Snippet: .. After an incubation of 48 h, the transfected cells were fixed with 4% paraformaldehyde in PBS for 15 min, permeabilized with 0.2% Triton X-100 for 10 min, blocked with 4% BSA in PBS for 1 h at 37°C, and then incubated with mouse primary anti-Myc antibodies (1 μg/ml) (Roche) for 1 h at 37°C. .. After three washes with PBS containing 0.2% Triton X-100, the cells were incubated with Alexa Fluor 488 -labeled anti-mouse secondary antibodies (Invitrogen) for 1 h at 37°C.

    Lysis:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Article Title: Molecular Characterization of the Host Defense Activity of the Barrier to Autointegration Factor against Vaccinia Virus ▿
    Article Snippet: .. Lysis was performed for 10 min on ice in lysis buffer (50 mM Tris [pH 7.4], 75 mM NaCl, 1 mM EDTA, 4% sucrose, and 0.5% Triton X-100) supplemented with fresh 1 mM phenylmethylsulfonyl fluoride (PMSF) and protease and phosphatase inhibitors (Complete Mini and PhosStop tablets; Roche), after which nuclei were removed following centrifugation at 800 × g . .. For DNA binding assays, 50 μl of native dsDNA cellulose beads (Amersham) was added to the lysates and incubated overnight at 4°C with end-over-end rotation.

    Western Blot:

    Article Title: Luteolin attenuates sepsis-induced myocardial injury by enhancing autophagy in mice
    Article Snippet: .. Western blot analysis Cardiac tissues or cardiomyocytes were harvested and homogenized with RIPA buffer containing protease inhibitor cocktail (Roche Diagnostics) on ice. .. After centrifugation at 12,000 × g for 15 min at 4°C, the total protein content of the supernatant was quantified using a bicinchoninic acid protein assay (Applygen Technologies, Inc.) and the supernatant was stored at −80°C until use.

    Staining:

    Article Title: The g5R (D250) Gene of African Swine Fever Virus Encodes a Nudix Hydrolase That Preferentially Degrades Diphosphoinositol Polyphosphates
    Article Snippet: .. At various times postinfection (7, 12, and 16 h) cells were fixed in 4% paraformaldehyde, permeabilized in 0.2% Triton X-100, and stained with rat anti-HA monoclonal antibody (1 in 600 dilution; Roche) and Alexa Fluor 568 goat anti-rat immunoglobulin G conjugate (1 in 800 dilution; Molecular Probes). .. Cells were visualized with a Leica TCS NT confocal microscope.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Roche fatty acid free bovine serum albumin bsa
    p21-dependent inhibition of <t>LPA-induced</t> cell proliferation by TGFβ. A . TGFβ inhibits LPA-afforded cell proliferation. MDA-MB-231 and Caov-3 cells in 6-well plates were incubated with LPA (10 µM) or vehicle <t>(BSA)</t> in the presence
    Fatty Acid Free Bovine Serum Albumin Bsa, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fatty acid free bovine serum albumin bsa/product/Roche
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    fatty acid free bovine serum albumin bsa - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    89
    Roche faf bsa
    Effects of <t>FAF-BSA</t> on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl
    Faf Bsa, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/faf bsa/product/Roche
    Average 89 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    faf bsa - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    Image Search Results


    p21-dependent inhibition of LPA-induced cell proliferation by TGFβ. A . TGFβ inhibits LPA-afforded cell proliferation. MDA-MB-231 and Caov-3 cells in 6-well plates were incubated with LPA (10 µM) or vehicle (BSA) in the presence

    Journal: Molecular Cancer Research

    Article Title: Lysophosphatidic acid-induced p21Waf1 expression mediates the cytostatic response of breast and ovarian cancer cells to transforming growth factor beta

    doi: 10.1158/1541-7786.MCR-11-0340

    Figure Lengend Snippet: p21-dependent inhibition of LPA-induced cell proliferation by TGFβ. A . TGFβ inhibits LPA-afforded cell proliferation. MDA-MB-231 and Caov-3 cells in 6-well plates were incubated with LPA (10 µM) or vehicle (BSA) in the presence

    Article Snippet: Prior to use, LPA and S1P were dissolved in PBS containing 0.5% fatty acid-free bovine serum albumin (BSA) obtained from Roche (Indianapolis, IN).

    Techniques: Inhibition, Multiple Displacement Amplification, Incubation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Journal:

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    doi: 10.1016/j.metabol.2007.09.015

    Figure Lengend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with KCl

    Article Snippet: In the initial series of studies we first confirmed the observations made previously by Straub and Sharp that a 4 hour incubation period in culture medium containing 100 μM FAF-BSA enhances glucose-induced insulin secretion.

    Techniques: Isolation

    Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Journal:

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    doi: 10.1016/j.metabol.2007.09.015

    Figure Lengend Snippet: Effects of FAF-BSA on insulin secretion in response to phorbol ester stimulation

    Article Snippet: In the initial series of studies we first confirmed the observations made previously by Straub and Sharp that a 4 hour incubation period in culture medium containing 100 μM FAF-BSA enhances glucose-induced insulin secretion.

    Techniques:

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Journal:

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    doi: 10.1016/j.metabol.2007.09.015

    Figure Lengend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused rat islets stimulated with 10 mM glucose

    Article Snippet: In the initial series of studies we first confirmed the observations made previously by Straub and Sharp that a 4 hour incubation period in culture medium containing 100 μM FAF-BSA enhances glucose-induced insulin secretion.

    Techniques: Isolation

    Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Journal:

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    doi: 10.1016/j.metabol.2007.09.015

    Figure Lengend Snippet: Effects of FAF-BSA on 3 H-inositol efflux from isolated, perifused rat islets stimulated with carbachol

    Article Snippet: In the initial series of studies we first confirmed the observations made previously by Straub and Sharp that a 4 hour incubation period in culture medium containing 100 μM FAF-BSA enhances glucose-induced insulin secretion.

    Techniques: Isolation

    Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Journal:

    Article Title: Enhanced Activation of Phospholipase C and Insulin Secretion from Islets Incubated in Fatty Acid-Free BSA

    doi: 10.1016/j.metabol.2007.09.015

    Figure Lengend Snippet: Effects of FAF-BSA on insulin secretion and 3 H-inositol efflux from isolated, perifused mouse islets

    Article Snippet: In the initial series of studies we first confirmed the observations made previously by Straub and Sharp that a 4 hour incubation period in culture medium containing 100 μM FAF-BSA enhances glucose-induced insulin secretion.

    Techniques: Isolation