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Omega Scientific Inc fetal bovine serum fbs
SGK3 promotes cell proliferation of AR-positive prostate cancer cells. A and B, LNCaP cells (A) or 22Rv1 cells (B) were transfected with the siRNA negative control (siNC) or 3 individual SGK3 siRNA duplexes (siSGK), respectively. Western blotting (insert) was performed to examine SGK3 level after the cells were transfected with siRNA for 72 hours. An MTT assay was performed to measure cell proliferation at the indicated time after siRNA transfection. Error bars represent SDs of 3 independent experiments. C, LNCaP/TetON/SGK3 cells cultured in phenol red–free RPMI <t>1640</t> medium with 2% CD-treated <t>FBS</t> were added with or without 100 ng/mL DOX. An MTT assay was performed to measure cell proliferation at the indicated time. D, LNCaP cells were transfected with the siRNA negative control or SGK3 siRNA and cultured in the phenol red–free RPMI 1640 medium with 5% CD-treated FBS in the presence or absence of DHT (1 nM). SGK3 protein levels were examined by Western blotting at 72 hours after siRNA transfection. The cell growth rate was measured by an MTT assay at the indicated time. **, P
Fetal Bovine Serum Fbs, supplied by Omega Scientific Inc, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "SGK3 Is an Androgen-Inducible Kinase Promoting Prostate Cancer Cell Proliferation Through Activation of p70 S6 Kinase and Up-Regulation of Cyclin D1"

Article Title: SGK3 Is an Androgen-Inducible Kinase Promoting Prostate Cancer Cell Proliferation Through Activation of p70 S6 Kinase and Up-Regulation of Cyclin D1

Journal: Molecular Endocrinology

doi: 10.1210/me.2013-1339

SGK3 promotes cell proliferation of AR-positive prostate cancer cells. A and B, LNCaP cells (A) or 22Rv1 cells (B) were transfected with the siRNA negative control (siNC) or 3 individual SGK3 siRNA duplexes (siSGK), respectively. Western blotting (insert) was performed to examine SGK3 level after the cells were transfected with siRNA for 72 hours. An MTT assay was performed to measure cell proliferation at the indicated time after siRNA transfection. Error bars represent SDs of 3 independent experiments. C, LNCaP/TetON/SGK3 cells cultured in phenol red–free RPMI 1640 medium with 2% CD-treated FBS were added with or without 100 ng/mL DOX. An MTT assay was performed to measure cell proliferation at the indicated time. D, LNCaP cells were transfected with the siRNA negative control or SGK3 siRNA and cultured in the phenol red–free RPMI 1640 medium with 5% CD-treated FBS in the presence or absence of DHT (1 nM). SGK3 protein levels were examined by Western blotting at 72 hours after siRNA transfection. The cell growth rate was measured by an MTT assay at the indicated time. **, P
Figure Legend Snippet: SGK3 promotes cell proliferation of AR-positive prostate cancer cells. A and B, LNCaP cells (A) or 22Rv1 cells (B) were transfected with the siRNA negative control (siNC) or 3 individual SGK3 siRNA duplexes (siSGK), respectively. Western blotting (insert) was performed to examine SGK3 level after the cells were transfected with siRNA for 72 hours. An MTT assay was performed to measure cell proliferation at the indicated time after siRNA transfection. Error bars represent SDs of 3 independent experiments. C, LNCaP/TetON/SGK3 cells cultured in phenol red–free RPMI 1640 medium with 2% CD-treated FBS were added with or without 100 ng/mL DOX. An MTT assay was performed to measure cell proliferation at the indicated time. D, LNCaP cells were transfected with the siRNA negative control or SGK3 siRNA and cultured in the phenol red–free RPMI 1640 medium with 5% CD-treated FBS in the presence or absence of DHT (1 nM). SGK3 protein levels were examined by Western blotting at 72 hours after siRNA transfection. The cell growth rate was measured by an MTT assay at the indicated time. **, P

Techniques Used: Transfection, Negative Control, Western Blot, MTT Assay, Cell Culture

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Article Snippet: .. Cells and viruses Vero, BHK21-15 and Raji-DC-SIGN-R cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and antibiotics (penicillin G and streptomycin) at 37°C in a 5% CO2 incubator. .. Strains from all five genotypes of DENV-1 included: TVP-2130 (genotype 1), 16007 (genotype 2), TVP-5175 (genotype 3), Western Pacific-74 (genotype 4) and 3146 Sri Lanka (genotype 5).

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Cell Culture:

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Article Snippet: .. Cells and viruses Vero, BHK21-15 and Raji-DC-SIGN-R cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and antibiotics (penicillin G and streptomycin) at 37°C in a 5% CO2 incubator. .. Strains from all five genotypes of DENV-1 included: TVP-2130 (genotype 1), 16007 (genotype 2), TVP-5175 (genotype 3), Western Pacific-74 (genotype 4) and 3146 Sri Lanka (genotype 5).

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Article Snippet: .. HEK293T cells were cultured with DMEM supplemented with 10% fetal bovine serum (FBS) (Omega Scientific) and 2% penicillin-streptomycin (Thermo Fisher Scientific). .. These cells were cultured at 37ºC, 5% CO2.

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    Omega Scientific Inc fbs
    Lipid raft and cholesterol content in T cells cultured in normal or <t>delipidated</t> medium. T cells isolated from naïve apoE-/- mice were cultured in culture medium supplemented with 10% delipidated <t>FBS</t> or 10% FBS medium for 24 hours with CD3/CD28 activation beads. More unesterified cholesterol contents (A) or lipid rafts (B) were observed in activated CD4 + and CD8 + T cells cultured in 10% FBS medium when compared to T cells cultured in 10% delipidated FBS medium. Experiments were repeated 4 times using T cells from 3 mice.
    Fbs, supplied by Omega Scientific Inc, used in various techniques. Bioz Stars score: 94/100, based on 196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Lipid raft and cholesterol content in T cells cultured in normal or delipidated medium. T cells isolated from naïve apoE-/- mice were cultured in culture medium supplemented with 10% delipidated FBS or 10% FBS medium for 24 hours with CD3/CD28 activation beads. More unesterified cholesterol contents (A) or lipid rafts (B) were observed in activated CD4 + and CD8 + T cells cultured in 10% FBS medium when compared to T cells cultured in 10% delipidated FBS medium. Experiments were repeated 4 times using T cells from 3 mice.

    Journal: PLoS ONE

    Article Title: Cholesterol Lowering Modulates T Cell Function In Vivo and In Vitro

    doi: 10.1371/journal.pone.0092095

    Figure Lengend Snippet: Lipid raft and cholesterol content in T cells cultured in normal or delipidated medium. T cells isolated from naïve apoE-/- mice were cultured in culture medium supplemented with 10% delipidated FBS or 10% FBS medium for 24 hours with CD3/CD28 activation beads. More unesterified cholesterol contents (A) or lipid rafts (B) were observed in activated CD4 + and CD8 + T cells cultured in 10% FBS medium when compared to T cells cultured in 10% delipidated FBS medium. Experiments were repeated 4 times using T cells from 3 mice.

    Article Snippet: Isolated T cells were loaded with 2.5 μM CFSE (Invitrogen) for 10 min and then seeded at 0.5×106 cells in 1 ml RPMI 1640 medium (invitrogen) supplemented with 10% FBS (Omega Scientific) or 10% delipidated FBS (Cocalico Biologicals Inc.), 1X Antibiotic-Antimycotic (Gibco) and 50 μM β-ME in 12 well plates.

    Techniques: Cell Culture, Isolation, Mouse Assay, Activation Assay

    T cell proliferation in normal or delipidated meidum using serum of the same lot. (A) T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4. (B) T cells isolated form naïve wild type mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4.

    Journal: PLoS ONE

    Article Title: Cholesterol Lowering Modulates T Cell Function In Vivo and In Vitro

    doi: 10.1371/journal.pone.0092095

    Figure Lengend Snippet: T cell proliferation in normal or delipidated meidum using serum of the same lot. (A) T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4. (B) T cells isolated form naïve wild type mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4.

    Article Snippet: Isolated T cells were loaded with 2.5 μM CFSE (Invitrogen) for 10 min and then seeded at 0.5×106 cells in 1 ml RPMI 1640 medium (invitrogen) supplemented with 10% FBS (Omega Scientific) or 10% delipidated FBS (Cocalico Biologicals Inc.), 1X Antibiotic-Antimycotic (Gibco) and 50 μM β-ME in 12 well plates.

    Techniques: Isolation, Mouse Assay, Cell Culture

    Western blot for pZap70 and IL-10 ELISA. (A) T cells isolated from naïve apoE-/- mice were cultured in 10% delipidated FBS medium or 10% FBS medium for 2 min with CD3/CD28 activation beads. Western blot analysis of the whole cell lysates revealed a higher expression of pZap-70 from T cells cultured in NM (medium supplemented with 10% FBS) when compared to that from T cells cultured in DM (medium supplemented with 10% delipidated FBS). Upper panel showed a representative Western blot and lower panel is the densitometric analysis from three experiments. N = 3 in each group. (B) IL-10 level was lower in delipidated medium of T cells after activation for 4 days with CD3/CD28 beads when compared to that in normal medium. N = 4.

    Journal: PLoS ONE

    Article Title: Cholesterol Lowering Modulates T Cell Function In Vivo and In Vitro

    doi: 10.1371/journal.pone.0092095

    Figure Lengend Snippet: Western blot for pZap70 and IL-10 ELISA. (A) T cells isolated from naïve apoE-/- mice were cultured in 10% delipidated FBS medium or 10% FBS medium for 2 min with CD3/CD28 activation beads. Western blot analysis of the whole cell lysates revealed a higher expression of pZap-70 from T cells cultured in NM (medium supplemented with 10% FBS) when compared to that from T cells cultured in DM (medium supplemented with 10% delipidated FBS). Upper panel showed a representative Western blot and lower panel is the densitometric analysis from three experiments. N = 3 in each group. (B) IL-10 level was lower in delipidated medium of T cells after activation for 4 days with CD3/CD28 beads when compared to that in normal medium. N = 4.

    Article Snippet: Isolated T cells were loaded with 2.5 μM CFSE (Invitrogen) for 10 min and then seeded at 0.5×106 cells in 1 ml RPMI 1640 medium (invitrogen) supplemented with 10% FBS (Omega Scientific) or 10% delipidated FBS (Cocalico Biologicals Inc.), 1X Antibiotic-Antimycotic (Gibco) and 50 μM β-ME in 12 well plates.

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Isolation, Mouse Assay, Cell Culture, Activation Assay, Expressing

    T cell proliferation in normal or delipidated medium. (A) T cells isolated from human peripheral blood were loaded with CFSE and cultured in culture medium supplemented with 10% delipidated FBS (delipidated) or 10% FBS (normal) for 4 days with CD3/CD28 activation beads. After activation, CD4 + and CD8 + T cells proliferated less when cultured in 10% delipidated FBS medium. Experiments were done in duplicates from the first sample, triplicates from the second sample and quadruplicates from the third sample using peripheral blood from 3 different human samples, hence n = 9 in each group. (B) Similarly CD4 + or CD8 + T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cultured in medium supplemented with 10% FBS. N = 6 in each group of CD4 + cells and n = 4 in each group of CD8 + cells.

    Journal: PLoS ONE

    Article Title: Cholesterol Lowering Modulates T Cell Function In Vivo and In Vitro

    doi: 10.1371/journal.pone.0092095

    Figure Lengend Snippet: T cell proliferation in normal or delipidated medium. (A) T cells isolated from human peripheral blood were loaded with CFSE and cultured in culture medium supplemented with 10% delipidated FBS (delipidated) or 10% FBS (normal) for 4 days with CD3/CD28 activation beads. After activation, CD4 + and CD8 + T cells proliferated less when cultured in 10% delipidated FBS medium. Experiments were done in duplicates from the first sample, triplicates from the second sample and quadruplicates from the third sample using peripheral blood from 3 different human samples, hence n = 9 in each group. (B) Similarly CD4 + or CD8 + T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cultured in medium supplemented with 10% FBS. N = 6 in each group of CD4 + cells and n = 4 in each group of CD8 + cells.

    Article Snippet: Isolated T cells were loaded with 2.5 μM CFSE (Invitrogen) for 10 min and then seeded at 0.5×106 cells in 1 ml RPMI 1640 medium (invitrogen) supplemented with 10% FBS (Omega Scientific) or 10% delipidated FBS (Cocalico Biologicals Inc.), 1X Antibiotic-Antimycotic (Gibco) and 50 μM β-ME in 12 well plates.

    Techniques: Isolation, Cell Culture, Activation Assay, Mouse Assay

    IGF-I–induced differentiation of multipotent neural progenitor cells. (A) Adult hippocampus-derived neural progenitor cells cultured in insulin-containing N2 media with 20 ng/ml FGF-2 (undifferentiated), 1 μM RA, and 1% FBS for 4 d (mixed), 1 μM RA and 5 μM forskolin for 4 d (neuronal), or 50 ng/ml leukemia inhibitory factor and 50 ng/ml BMP2 for 6 d (astrocytic). Cells were stained for markers for neurons (Tuj1), astrocytes (GFAP), or oligodendrocytes (RIP), and also with DAPI. Bar, 50 μm. (B) Differentiation of neural progenitor cells in insulin-free N2 media without IGF-I (control) or with IGF-I for 4 d. Insets show cells of typical oligodendrocyte morphology stained with RIP or MBP. Bar, 25 μm. (C) Quantification of cells in either proliferating or differentiating conditions. (D) Quantification of cultures grown in insulin-free N2 media (control) or treated with 500 ng/ml IGF-I, IGF-II, or insulin after 4 d. (E) Quantification of cells in response to different doses of IGF-I (20–500 ng/ml) in 4-d cultures. All data shown are from at least three experiments in parallel cultures with error bars representing SDs. Significant differences are indicated with an asterisk (P

    Journal: The Journal of Cell Biology

    Article Title: IGF-I instructs multipotent adult neural progenitor cells to become oligodendrocytes

    doi: 10.1083/jcb.200308101

    Figure Lengend Snippet: IGF-I–induced differentiation of multipotent neural progenitor cells. (A) Adult hippocampus-derived neural progenitor cells cultured in insulin-containing N2 media with 20 ng/ml FGF-2 (undifferentiated), 1 μM RA, and 1% FBS for 4 d (mixed), 1 μM RA and 5 μM forskolin for 4 d (neuronal), or 50 ng/ml leukemia inhibitory factor and 50 ng/ml BMP2 for 6 d (astrocytic). Cells were stained for markers for neurons (Tuj1), astrocytes (GFAP), or oligodendrocytes (RIP), and also with DAPI. Bar, 50 μm. (B) Differentiation of neural progenitor cells in insulin-free N2 media without IGF-I (control) or with IGF-I for 4 d. Insets show cells of typical oligodendrocyte morphology stained with RIP or MBP. Bar, 25 μm. (C) Quantification of cells in either proliferating or differentiating conditions. (D) Quantification of cultures grown in insulin-free N2 media (control) or treated with 500 ng/ml IGF-I, IGF-II, or insulin after 4 d. (E) Quantification of cells in response to different doses of IGF-I (20–500 ng/ml) in 4-d cultures. All data shown are from at least three experiments in parallel cultures with error bars representing SDs. Significant differences are indicated with an asterisk (P

    Article Snippet: Differentiation conditions were either N2 medium with 1 μM RA (Sigma-Aldrich) and 1% FBS (Omega Scientific) for 4 d (mixed); 1 μM RA and 5 μM forskolin (Sigma-Aldrich) for 4 d (neuronal), or 50 ng/ml leukemia inhibitory factor (CHEMICON International, Inc.) and 50 ng/ml BMP2 (R & D Systems) for 6 d (astrocytic; ).

    Techniques: Derivative Assay, Cell Culture, Staining

    ATF6(N) associates with SREBP2(N). ( A ) Cell lysates collected from HEK293 cells transfected with pCI-Flag-ATF6(N) were incubated with GST or GST-SREBP2(N) glutathione beads for GST pull-down assays. The precipitates were separated by SDS–PAGE and immunoblotted with anti-Flag. The input control was 10% of the cell lysates. ( B ) HEK293 cells were transiently transfected with pCMV5-HA-SREBP2(N) and pCI-Flag-ATF6(N). HA-SREBP2(N) was immunoprecipitated from the cell lysates by anti-HA, and the associated ATF6(N) was revealed by immunoblotting with anti-Flag. In control experiments, rabbit IgG was used in immunoprecipitation, and the input was 10% of the cell lysates. Shown at the bottom is the anti-HA immunoblotting demonstrating the expression of HA-SREBP2(N). ( C ) HepG2 cells were transiently transfected with pCGN-HA-ATF6. At 1 day after transfection, the media were changed to normal growth media or glucose-deprived media for the indicated times. HA-ATF6 was immunoprecipitated from the cell lysates by anti-HA, and the endogenous SREBP2 associated with HA-ATF6 was examined by immunoblotting with anti-SREBP2 pAb. The membrane was stripped and reprobed with mouse anti-HA antibody to reveal the expression of the ATF6 precursor and its cleavage in response to glucose deprivation (bottom panel). ( D ) HEK293 cells were subjected to glucose-deficient media containing 10% dialyzed FBS for 1 h or kept in high glucose media for the same period of time. Endogenous SREBP2 was immunoprecipitated from the cell lysates by anti-SREBP2 pAb. The associated ATF6 was detected by immunoblotting with anti-ATF6 pAb. In control experiments, the input was 10% of the cell lysates, and goat IgG was used in an immunoprecipitation control. The membrane was stripped and reprobed with anti-SREBP2 pAb (bottom panel).

    Journal: The EMBO Journal

    Article Title: ATF6 modulates SREBP2-mediated lipogenesis

    doi: 10.1038/sj.emboj.7600106

    Figure Lengend Snippet: ATF6(N) associates with SREBP2(N). ( A ) Cell lysates collected from HEK293 cells transfected with pCI-Flag-ATF6(N) were incubated with GST or GST-SREBP2(N) glutathione beads for GST pull-down assays. The precipitates were separated by SDS–PAGE and immunoblotted with anti-Flag. The input control was 10% of the cell lysates. ( B ) HEK293 cells were transiently transfected with pCMV5-HA-SREBP2(N) and pCI-Flag-ATF6(N). HA-SREBP2(N) was immunoprecipitated from the cell lysates by anti-HA, and the associated ATF6(N) was revealed by immunoblotting with anti-Flag. In control experiments, rabbit IgG was used in immunoprecipitation, and the input was 10% of the cell lysates. Shown at the bottom is the anti-HA immunoblotting demonstrating the expression of HA-SREBP2(N). ( C ) HepG2 cells were transiently transfected with pCGN-HA-ATF6. At 1 day after transfection, the media were changed to normal growth media or glucose-deprived media for the indicated times. HA-ATF6 was immunoprecipitated from the cell lysates by anti-HA, and the endogenous SREBP2 associated with HA-ATF6 was examined by immunoblotting with anti-SREBP2 pAb. The membrane was stripped and reprobed with mouse anti-HA antibody to reveal the expression of the ATF6 precursor and its cleavage in response to glucose deprivation (bottom panel). ( D ) HEK293 cells were subjected to glucose-deficient media containing 10% dialyzed FBS for 1 h or kept in high glucose media for the same period of time. Endogenous SREBP2 was immunoprecipitated from the cell lysates by anti-SREBP2 pAb. The associated ATF6 was detected by immunoblotting with anti-ATF6 pAb. In control experiments, the input was 10% of the cell lysates, and goat IgG was used in an immunoprecipitation control. The membrane was stripped and reprobed with anti-SREBP2 pAb (bottom panel).

    Article Snippet: For glucose deprivation, HepG2 or HEK293 cells were subjected to glucose-deficient DMEM (Gibco/BRL) supplemented with 10% dialyzed FBS (Omega Scientific, Tarzana, CA) for the time periods indicated.

    Techniques: Transfection, Incubation, SDS Page, Immunoprecipitation, Expressing

    Glucose deprivation suppresses SREBP2-mediated but activates ATF6-mediated transcription. ( A ) HepG2 and HEK293 cells cultured in high glucose (27.5 mM) DMEM with 10% FBS were subjected to glucose-deficient DMEM with 10% dialyzed FBS for 0, 3, and 6 h. Total RNA was isolated for Northern blotting probed with HMG-CoA reductase, HMG-CoA synthase, squalene synthase, LDLR, BiP/grp78, GADD153, and β-actin cDNA. ( B ) HepG2 cells in 12-well plates were transiently transfected with 4 × SRE-Luc, LDLR-Luc, 5 × ATF6-Luc, or BiP/grp78-Luc together with renilla-Luc for 24 h. The transfected cells were then cultured in DMEM containing 0, 5.5, 27.5 mM glucose, or 10 mM lactate for 12 h. In parallel control experiments, 300 nM Tg or 10% LDS was included in the DMEM containing 27.5 mM glucose. The cells were then lysed for the measurement of firefly and renilla luciferase activity. The relative luciferase activity is defined as the firefly luciferase activity normalized to that of renilla luciferase and that of cells under 27.5 mM glucose is set as 1.

    Journal: The EMBO Journal

    Article Title: ATF6 modulates SREBP2-mediated lipogenesis

    doi: 10.1038/sj.emboj.7600106

    Figure Lengend Snippet: Glucose deprivation suppresses SREBP2-mediated but activates ATF6-mediated transcription. ( A ) HepG2 and HEK293 cells cultured in high glucose (27.5 mM) DMEM with 10% FBS were subjected to glucose-deficient DMEM with 10% dialyzed FBS for 0, 3, and 6 h. Total RNA was isolated for Northern blotting probed with HMG-CoA reductase, HMG-CoA synthase, squalene synthase, LDLR, BiP/grp78, GADD153, and β-actin cDNA. ( B ) HepG2 cells in 12-well plates were transiently transfected with 4 × SRE-Luc, LDLR-Luc, 5 × ATF6-Luc, or BiP/grp78-Luc together with renilla-Luc for 24 h. The transfected cells were then cultured in DMEM containing 0, 5.5, 27.5 mM glucose, or 10 mM lactate for 12 h. In parallel control experiments, 300 nM Tg or 10% LDS was included in the DMEM containing 27.5 mM glucose. The cells were then lysed for the measurement of firefly and renilla luciferase activity. The relative luciferase activity is defined as the firefly luciferase activity normalized to that of renilla luciferase and that of cells under 27.5 mM glucose is set as 1.

    Article Snippet: For glucose deprivation, HepG2 or HEK293 cells were subjected to glucose-deficient DMEM (Gibco/BRL) supplemented with 10% dialyzed FBS (Omega Scientific, Tarzana, CA) for the time periods indicated.

    Techniques: Cell Culture, Isolation, Northern Blot, Transfection, Luciferase, Activity Assay

    SGK3 promotes cell proliferation of AR-positive prostate cancer cells. A and B, LNCaP cells (A) or 22Rv1 cells (B) were transfected with the siRNA negative control (siNC) or 3 individual SGK3 siRNA duplexes (siSGK), respectively. Western blotting (insert) was performed to examine SGK3 level after the cells were transfected with siRNA for 72 hours. An MTT assay was performed to measure cell proliferation at the indicated time after siRNA transfection. Error bars represent SDs of 3 independent experiments. C, LNCaP/TetON/SGK3 cells cultured in phenol red–free RPMI 1640 medium with 2% CD-treated FBS were added with or without 100 ng/mL DOX. An MTT assay was performed to measure cell proliferation at the indicated time. D, LNCaP cells were transfected with the siRNA negative control or SGK3 siRNA and cultured in the phenol red–free RPMI 1640 medium with 5% CD-treated FBS in the presence or absence of DHT (1 nM). SGK3 protein levels were examined by Western blotting at 72 hours after siRNA transfection. The cell growth rate was measured by an MTT assay at the indicated time. **, P

    Journal: Molecular Endocrinology

    Article Title: SGK3 Is an Androgen-Inducible Kinase Promoting Prostate Cancer Cell Proliferation Through Activation of p70 S6 Kinase and Up-Regulation of Cyclin D1

    doi: 10.1210/me.2013-1339

    Figure Lengend Snippet: SGK3 promotes cell proliferation of AR-positive prostate cancer cells. A and B, LNCaP cells (A) or 22Rv1 cells (B) were transfected with the siRNA negative control (siNC) or 3 individual SGK3 siRNA duplexes (siSGK), respectively. Western blotting (insert) was performed to examine SGK3 level after the cells were transfected with siRNA for 72 hours. An MTT assay was performed to measure cell proliferation at the indicated time after siRNA transfection. Error bars represent SDs of 3 independent experiments. C, LNCaP/TetON/SGK3 cells cultured in phenol red–free RPMI 1640 medium with 2% CD-treated FBS were added with or without 100 ng/mL DOX. An MTT assay was performed to measure cell proliferation at the indicated time. D, LNCaP cells were transfected with the siRNA negative control or SGK3 siRNA and cultured in the phenol red–free RPMI 1640 medium with 5% CD-treated FBS in the presence or absence of DHT (1 nM). SGK3 protein levels were examined by Western blotting at 72 hours after siRNA transfection. The cell growth rate was measured by an MTT assay at the indicated time. **, P

    Article Snippet: Human prostate cancer cell lines LNCaP, 22Rv1, and PC3 and human benign prostatic hyperplasia (BPH) epithelial cell line BPH-1 were propagated in RPMI 1640 medium (HyClone Laboratories, Inc) supplemented with 2 mM l -glutamine, 10% fetal bovine serum (FBS) (Omega Scientific), and 100 U/mL penicillin-streptomycin.

    Techniques: Transfection, Negative Control, Western Blot, MTT Assay, Cell Culture