dmem  (Lonza)


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    Name:
    DMEM with Glucose without L Glutamine
    Description:
    Dulbecco s Modified Eagle Medium DMEM with 4 5 g L Glucose without L Glutamine 500 mL
    Catalog Number:
    be12-614f
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    None
    Category:
    Culture Media and Reagents
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    Structured Review

    Lonza dmem
    Dulbecco s Modified Eagle Medium DMEM with 4 5 g L Glucose without L Glutamine 500 mL
    https://www.bioz.com/result/dmem/product/Lonza
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dmem - by Bioz Stars, 2021-03
    97/100 stars

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    Related Articles

    Multiple Displacement Amplification:

    Article Title: CRP2, a new invadopodia actin bundling factor critically promotes breast cancer cell invasion and metastasis
    Article Snippet: MCF-7-snail 6SA were maintained in DMEM:F12 (1:1; Lonza) supplemented with 1mM Sodium Pyruvate (Lonza) and 500 μg/ml G418 (Sigma-Aldrich). .. MCF-7, MDA-MB-231, MDA-MB-231-luc and 1001 cells were maintained in DMEM high glucose with L-glutamine medium (Lonza). ..

    Cell Culture:

    Article Title: Plasma Membrane Repair Is Regulated Extracellularly by Proteases Released from Lysosomes
    Article Snippet: .. Cell Culture HeLa (ATCC) and NRK (ATCC) cells were cultivated at 37°C/5% C02 in high glucose DMEM (Dulbecco’s Modified Eagle’s Medium) (Lonza) containing 10% FBS (fetal bovine serum) and antibiotics (penicillin/streptomycin) (Gemini). .. Cells were plated 48 h before the experiments to obtain sub-confluent cell monolayers in 35 mm glass-bottom microwell dishes (MatTek), 35 or 100 mm plastic dishes (Cellstar), or glass coverslips.

    Article Title: Modelling the endothelial blood-CNS barriers: a method for the production of robust in vitro models of the rat blood-brain barrier and blood-spinal cord barrier
    Article Snippet: We investigated whether commercially available speciality endothelial cell culture reagents could influence both the quantity of endothelial cells recovered and the quality of the rat in vitro barriers generated by this passaging technique. .. Following initial plating in DMEM with MVGS supplement, we passaged the RBECs onto collagen I/fibronectin-coated cell culture inserts and compared two media formulations in both the top well and bottom well: (a) DMEM with MVGS, and (b) EBM-2 microvascular endothelial cell media with the EGM-2 BulletKit without VEGF (Lonza). .. The endothelial cells were co-cultured with mixed glia plated into the bottom chamber of the dish, as the role of astrocytes in inducing barrier phenotype in primary brain endothelial cells in vitro is well validated [ , , , ].

    Article Title: Upregulation of skeletal muscle PGC-1α through the elevation of cyclic AMP levels by Cyanidin-3-glucoside enhances exercise performance
    Article Snippet: .. Cell culture and differentiation The mouse C2C12 myoblasts (ATCC, USA) were cultured in DMEM supplemented with 10% foetal bovine serum and 1% penicillin (5000 μg/ml)–streptomycin (5000 IU/ml) (Lonza, Tokyo, Japan). .. Normal human skeletal muscle myoblasts (Lonza) were cultured in a SkGM-2 Bullet Kit (Lonza) at 37 °C in a humidified atmosphere of 5% CO2 .

    Article Title: Expansion of bone marrow‐derived human mesenchymal stem/stromal cells (hMSCs) using a two‐phase liquid/liquid system
    Article Snippet: .. The cells were cultured according to previously determined protocols., , Briefly, the cells were seeded at 5000 cells cm−2 and grown on tissue culture plastic (TCPS) (NUNC, ThermoFisher, UK) in DMEM supplemented with 10% (v/v) foetal bovine serum and 2 mmol L−1 ultra‐glutamine (Lonza, UK). ..

    Article Title: Distinct phosphorylation sites on the ghrelin receptor, GHSR1a, establish a code that determines the functions of ß-arrestins
    Article Snippet: .. Cell culture The human embryonic kidney (HEK) 293 wild-type (WT) cell line was maintained in DMEM (Lonza; Basilea, CH) supplemented with fetal bovine serum [FBS, 10% (v/v)], and 100 U/mL penicillin-streptomycin solution (100U/mL). .. The wild-type (WT), the ß-arrestin 1 knockout (ß-arrestin-1−/− ) and the ß-arrestin 2 knockout (ß-arrestin-2−/− ) murine embryonic fibroblast (MEF) cells, provided by Prof. Robert J. Lefkowitz (Duke University Medical Center, Durham, NC, US), were maintained in DMEM supplemented with 10% (v/v) FBS and 100 U/mL penicillin-streptomycin solution.

    Modification:

    Article Title: Plasma Membrane Repair Is Regulated Extracellularly by Proteases Released from Lysosomes
    Article Snippet: .. Cell Culture HeLa (ATCC) and NRK (ATCC) cells were cultivated at 37°C/5% C02 in high glucose DMEM (Dulbecco’s Modified Eagle’s Medium) (Lonza) containing 10% FBS (fetal bovine serum) and antibiotics (penicillin/streptomycin) (Gemini). .. Cells were plated 48 h before the experiments to obtain sub-confluent cell monolayers in 35 mm glass-bottom microwell dishes (MatTek), 35 or 100 mm plastic dishes (Cellstar), or glass coverslips.

    Negative Control:

    Article Title: Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC)
    Article Snippet: .. We used DMEM as a negative control and EGM-2 (Endothelial Cell Growth Medium; Lonza) as a positive control. ..

    Positive Control:

    Article Title: Sonic hedgehog (SHH) signaling improves the angiogenic potential of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSC)
    Article Snippet: .. We used DMEM as a negative control and EGM-2 (Endothelial Cell Growth Medium; Lonza) as a positive control. ..

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  • 86
    Lonza dmem f12
    UC-MSC-derived neurospheres can be generated using one-step induction. (a) UC-MSC-derived neurosphere formation at different timepoints: 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after induction. (b) Representative phase contrast images (100x) of UC-MSC-derived neurospheres formed under different regimens, including (1) EGF, bFGF only regimen: EGF and bFGF, both at 20 ng/ml; (2) complete NB (neurobasal) medium regimen (EGF and bFGF, both at 20 ng/ml, with N2 and B27 supplements); (3) half regimen: EGF and bFGF, both at 10 ng/ml, with N2 and B27 supplements; (4) EGF only regimen: EGF 20 ng/ml, no bFGF, with N2 and B27 supplements; (5) bFGF only regimen: bFGF 20 ng/ml, no EGF, with N2 and B27 supplements; (6) N2 only regimen: <t>DMEM/F12</t> with N2 supplement only, no EGF nor bFGF; and (7) B27 only regimen: DMEM/F12 with B27 supplement only, no EGF nor bFGF. Enlarged image of the black-boxed region denotes the arbitrary classification of large-, medium-, and small-sized neurospheres. Blue arrows denote irregular cell clumps. (c) Statistical analysis of large-, medium-, and small-sized neurospheres produced under different induction regimens. Neurospheres were scored in 6 random fields under the microscope at 100x magnification. n = 3. ∗∗ p
    Dmem F12, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dmem f12/product/Lonza
    Average 86 stars, based on 1 article reviews
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    94
    Lonza serum free dmem
    WNT Treatment Induces AAK1-, PIP2-Dependent Phosphorylation of AP2M1 (A) Western blot analysis of HT1080 cells treated with WNT3A CM for indicated times. (B) Western blot analysis of HT1080 cells transfected with either AAK1 or control siRNA for 72 hr. Cells were then treated with rhWNT3A (200 ng/mL) for 15 min, then the medium was changed for fresh, complete <t>DMEM,</t> and cells were incubated for 9 hr. (C) Western blot analysis of pAP2M1 levels in <t>HEK293T</t> cells treated with either rhWNT3A (3A) or rhWNT5A (5A) (200 ng/mL) for 9 hr. The box-and-whisker plot represents pAP2M1/AP2M1 expression from four biological replicates quantified by LiCOR software. (D) Western blot analysis of HT1080 cells treated with either CHIR99021 compound (1 μM) or rhWNT3A (200 ng/mL) for the indicated time. (E) HEK293T cells were transfected with FLAG-dnTCF7L2 for 24 hr. Cells were then treated with WNT3A for 15 min, then the medium was changed for fresh, complete DMEM, incubated for 9 hr, and analyzed using western blot. (F) Western blot analysis of HEK293T WT, DVL TKO clone #4, DVL TKO clone #6, and LRP5/6 DKO cells treated with WNT3A CM for 9 hr. (G) Western blot analysis of HT1080 cells treated with WNT3A CM for 15 mins, then the medium was changed for complete DMEM containing either carbachol (50 μM) or neomycin (100 μM). Cells were incubated for 9 hr prior to cell harvest. (H) Western blot analysis of HT1080 cells transfected with siRNA against PI4K2α for 72 hr and pulsed with WNT3A CM for 15 mins and then incubated for 9 hr. All panels are representative of biological triplicates, unless otherwise noted. For complete statistics, see STAR Methods . See also Figure S4 .
    Serum Free Dmem, supplied by Lonza, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/serum free dmem/product/Lonza
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    dmem  (Lonza)
    99
    Lonza dmem
    mTOR inhibition leads to SRC activation. ( A ) <t>U87MG</t> cells were cultured in complete <t>DMEM</t> (CTR) or aminoacid- and serum- free medium (EBSS) or in DMEM in the presence of 250 nM Torin1 or 100 nM AZD8055 for 18 h. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. ( B ) U87MG cells were cultured in DMEM in the presence of Torin1 pre-incubating or not with 20 µM PP2 inhibitor for the indicated time points. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. The graphs represent the mean ± SD of three different experiments. Statistical significance: ** p
    Dmem, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dmem/product/Lonza
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    99/100 stars
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    Image Search Results


    UC-MSC-derived neurospheres can be generated using one-step induction. (a) UC-MSC-derived neurosphere formation at different timepoints: 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after induction. (b) Representative phase contrast images (100x) of UC-MSC-derived neurospheres formed under different regimens, including (1) EGF, bFGF only regimen: EGF and bFGF, both at 20 ng/ml; (2) complete NB (neurobasal) medium regimen (EGF and bFGF, both at 20 ng/ml, with N2 and B27 supplements); (3) half regimen: EGF and bFGF, both at 10 ng/ml, with N2 and B27 supplements; (4) EGF only regimen: EGF 20 ng/ml, no bFGF, with N2 and B27 supplements; (5) bFGF only regimen: bFGF 20 ng/ml, no EGF, with N2 and B27 supplements; (6) N2 only regimen: DMEM/F12 with N2 supplement only, no EGF nor bFGF; and (7) B27 only regimen: DMEM/F12 with B27 supplement only, no EGF nor bFGF. Enlarged image of the black-boxed region denotes the arbitrary classification of large-, medium-, and small-sized neurospheres. Blue arrows denote irregular cell clumps. (c) Statistical analysis of large-, medium-, and small-sized neurospheres produced under different induction regimens. Neurospheres were scored in 6 random fields under the microscope at 100x magnification. n = 3. ∗∗ p

    Journal: Stem Cells International

    Article Title: Efficient One-Step Induction of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Produces MSC-Derived Neurospheres (MSC-NS) with Unique Transcriptional Profile and Enhanced Neurogenic and Angiogenic Secretomes

    doi: 10.1155/2019/9208173

    Figure Lengend Snippet: UC-MSC-derived neurospheres can be generated using one-step induction. (a) UC-MSC-derived neurosphere formation at different timepoints: 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after induction. (b) Representative phase contrast images (100x) of UC-MSC-derived neurospheres formed under different regimens, including (1) EGF, bFGF only regimen: EGF and bFGF, both at 20 ng/ml; (2) complete NB (neurobasal) medium regimen (EGF and bFGF, both at 20 ng/ml, with N2 and B27 supplements); (3) half regimen: EGF and bFGF, both at 10 ng/ml, with N2 and B27 supplements; (4) EGF only regimen: EGF 20 ng/ml, no bFGF, with N2 and B27 supplements; (5) bFGF only regimen: bFGF 20 ng/ml, no EGF, with N2 and B27 supplements; (6) N2 only regimen: DMEM/F12 with N2 supplement only, no EGF nor bFGF; and (7) B27 only regimen: DMEM/F12 with B27 supplement only, no EGF nor bFGF. Enlarged image of the black-boxed region denotes the arbitrary classification of large-, medium-, and small-sized neurospheres. Blue arrows denote irregular cell clumps. (c) Statistical analysis of large-, medium-, and small-sized neurospheres produced under different induction regimens. Neurospheres were scored in 6 random fields under the microscope at 100x magnification. n = 3. ∗∗ p

    Article Snippet: We tested several different conditions: (1) complete neurobasal medium which is comprised of DMEM/F12, EGF 20 ng/ml, bFGF 20 ng/ml, and N2 and B27 supplements; (2) EGF and bFGF only: DMEM/F12, EGF 20 ng/ml, and bFGF 20 ng/ml, without N2 and B27 supplements; (3) half regimen: DMEM/F12, EGF 10 ng/ml, and bFGF 10 ng/ml, with N2 and B27 supplements; (4) EGF only regimen: DMEM/F12, EGF 20 ng/ml, and no bFGF, with N2 and B27 supplements; (5) bFGF only regimen: DMEM/F12, bFGF 20 ng/ml, and no EGF, with N2 and B27 supplements; (6) N2 only regimen: DMEM/F12 with N2 supplement only, no EGF or bFGF; and (7) B27 only regimen: DMEM/F12 with B27 supplement only, no EGF or bFGF.

    Techniques: Derivative Assay, Generated, Produced, Microscopy

    WNT Treatment Induces AAK1-, PIP2-Dependent Phosphorylation of AP2M1 (A) Western blot analysis of HT1080 cells treated with WNT3A CM for indicated times. (B) Western blot analysis of HT1080 cells transfected with either AAK1 or control siRNA for 72 hr. Cells were then treated with rhWNT3A (200 ng/mL) for 15 min, then the medium was changed for fresh, complete DMEM, and cells were incubated for 9 hr. (C) Western blot analysis of pAP2M1 levels in HEK293T cells treated with either rhWNT3A (3A) or rhWNT5A (5A) (200 ng/mL) for 9 hr. The box-and-whisker plot represents pAP2M1/AP2M1 expression from four biological replicates quantified by LiCOR software. (D) Western blot analysis of HT1080 cells treated with either CHIR99021 compound (1 μM) or rhWNT3A (200 ng/mL) for the indicated time. (E) HEK293T cells were transfected with FLAG-dnTCF7L2 for 24 hr. Cells were then treated with WNT3A for 15 min, then the medium was changed for fresh, complete DMEM, incubated for 9 hr, and analyzed using western blot. (F) Western blot analysis of HEK293T WT, DVL TKO clone #4, DVL TKO clone #6, and LRP5/6 DKO cells treated with WNT3A CM for 9 hr. (G) Western blot analysis of HT1080 cells treated with WNT3A CM for 15 mins, then the medium was changed for complete DMEM containing either carbachol (50 μM) or neomycin (100 μM). Cells were incubated for 9 hr prior to cell harvest. (H) Western blot analysis of HT1080 cells transfected with siRNA against PI4K2α for 72 hr and pulsed with WNT3A CM for 15 mins and then incubated for 9 hr. All panels are representative of biological triplicates, unless otherwise noted. For complete statistics, see STAR Methods . See also Figure S4 .

    Journal: Cell Reports

    Article Title: WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop

    doi: 10.1016/j.celrep.2018.12.023

    Figure Lengend Snippet: WNT Treatment Induces AAK1-, PIP2-Dependent Phosphorylation of AP2M1 (A) Western blot analysis of HT1080 cells treated with WNT3A CM for indicated times. (B) Western blot analysis of HT1080 cells transfected with either AAK1 or control siRNA for 72 hr. Cells were then treated with rhWNT3A (200 ng/mL) for 15 min, then the medium was changed for fresh, complete DMEM, and cells were incubated for 9 hr. (C) Western blot analysis of pAP2M1 levels in HEK293T cells treated with either rhWNT3A (3A) or rhWNT5A (5A) (200 ng/mL) for 9 hr. The box-and-whisker plot represents pAP2M1/AP2M1 expression from four biological replicates quantified by LiCOR software. (D) Western blot analysis of HT1080 cells treated with either CHIR99021 compound (1 μM) or rhWNT3A (200 ng/mL) for the indicated time. (E) HEK293T cells were transfected with FLAG-dnTCF7L2 for 24 hr. Cells were then treated with WNT3A for 15 min, then the medium was changed for fresh, complete DMEM, incubated for 9 hr, and analyzed using western blot. (F) Western blot analysis of HEK293T WT, DVL TKO clone #4, DVL TKO clone #6, and LRP5/6 DKO cells treated with WNT3A CM for 9 hr. (G) Western blot analysis of HT1080 cells treated with WNT3A CM for 15 mins, then the medium was changed for complete DMEM containing either carbachol (50 μM) or neomycin (100 μM). Cells were incubated for 9 hr prior to cell harvest. (H) Western blot analysis of HT1080 cells transfected with siRNA against PI4K2α for 72 hr and pulsed with WNT3A CM for 15 mins and then incubated for 9 hr. All panels are representative of biological triplicates, unless otherwise noted. For complete statistics, see STAR Methods . See also Figure S4 .

    Article Snippet: The transfection complexes were then mixed with HEK293T cells in a 100 mm dish at 50%–70% confluence in serum-free DMEM (Lonza), followed by incubation in a humidified, 37°C, 5% CO2 incubator.

    Techniques: Western Blot, Transfection, Incubation, Whisker Assay, Expressing, Software

    mTOR inhibition leads to SRC activation. ( A ) U87MG cells were cultured in complete DMEM (CTR) or aminoacid- and serum- free medium (EBSS) or in DMEM in the presence of 250 nM Torin1 or 100 nM AZD8055 for 18 h. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. ( B ) U87MG cells were cultured in DMEM in the presence of Torin1 pre-incubating or not with 20 µM PP2 inhibitor for the indicated time points. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. The graphs represent the mean ± SD of three different experiments. Statistical significance: ** p

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: mTOR inhibition leads to SRC activation. ( A ) U87MG cells were cultured in complete DMEM (CTR) or aminoacid- and serum- free medium (EBSS) or in DMEM in the presence of 250 nM Torin1 or 100 nM AZD8055 for 18 h. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. ( B ) U87MG cells were cultured in DMEM in the presence of Torin1 pre-incubating or not with 20 µM PP2 inhibitor for the indicated time points. Western blot analysis was performed by using specific antibodies for the total (SRC) and the phosphorylated (Y416) form of SRC (P-Src). β-ACTIN was used as loading control. The graphs represent the mean ± SD of three different experiments. Statistical significance: ** p

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Inhibition, Activation Assay, Cell Culture, Western Blot

    The mitogen activated protein (MAPK)/ERK pathway is down-regulated upon mTOR inhibition in GBM cells. ( A , B ) U87MG, GL15 were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at 2 h and 4 h ( A ) and at 24 h and 48 h ( B ). Western blot analysis was performed by using specific antibodies for ERK1/2 and P-ERK1/2. HSP90 was used as loading control. ( C ) GH2 cells were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at the indicated time points. Western blot analysis was performed by using specific antibodies for ERK1/2 and P-ERK1/2. HSP90 was used as loading control. The graph represents the mean ± SD of three different experiments. Statistical significance: * p

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: The mitogen activated protein (MAPK)/ERK pathway is down-regulated upon mTOR inhibition in GBM cells. ( A , B ) U87MG, GL15 were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at 2 h and 4 h ( A ) and at 24 h and 48 h ( B ). Western blot analysis was performed by using specific antibodies for ERK1/2 and P-ERK1/2. HSP90 was used as loading control. ( C ) GH2 cells were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at the indicated time points. Western blot analysis was performed by using specific antibodies for ERK1/2 and P-ERK1/2. HSP90 was used as loading control. The graph represents the mean ± SD of three different experiments. Statistical significance: * p

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Inhibition, Cell Culture, Western Blot

    mTOR inhibition leads to a reduction of EGFR expression in GBM cells. ( A , B ) U87MG cells were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at the indicated time points. Western blot analyses were performed by using a specific antibody for EGFR. HSP90 was used as loading control. ( C ) Real time experiments were performed on U87MG and GL15 cells stimulated or not with 250 nM Torin1 for 24 h. GAPDH was used as internal control. The graph represents the mean ± SD of three different experiments: *** p

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: mTOR inhibition leads to a reduction of EGFR expression in GBM cells. ( A , B ) U87MG cells were cultured in complete DMEM medium in the presence (TORIN1) or absence (CTR) of 250 nM Torin1 and analysed at the indicated time points. Western blot analyses were performed by using a specific antibody for EGFR. HSP90 was used as loading control. ( C ) Real time experiments were performed on U87MG and GL15 cells stimulated or not with 250 nM Torin1 for 24 h. GAPDH was used as internal control. The graph represents the mean ± SD of three different experiments: *** p

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Inhibition, Expressing, Cell Culture, Western Blot

    Epidermal Growth Factor receptor (EGFR) internalises into Glioblastoma multiforme (GBM) cells upon mTOR inhibition. ( A ) U87MG (upper panels), GL15 (middle panels) and primary GH2 cells (lower panels) were cultured in complete medium (DMEM) (CTR) or in DMEM in the presence of 250 nM Torin1 for 18 h. Immunocytochemistry and confocal analysis for EGFR localisation (red) were then performed. Hoechst 33342 was used to stain nuclei (blue). Scale bar, 30 μM. Western blot analysis of P-p70S6K and p70S6K was performed to check mTOR pathway inhibition by Torin1. β-actin was used as loading control. The blots are representative of three independent experiments. ( B ) Immunocytochemistry and confocal analysis for EGFR localisation (red) were performed in U87MG cells, upon 6 h and 24 h Torin1 treatment in the presence or absence of 100 μM Dynasore. Scale bar, 30 μM. A 4× magnification is shown for the right panels representing cells treated with Torin1 plus Dynasore. ( C ) U87MG cells were incubated with 0.5 mg/mL Rhodamine Dextran for the indicated times and its uptake within the cells analysed by image capturing at the confocal microscope. Scale bar, 30 μM. Fluorescence quantification of Dextran uptake at 120′ is shown in the right panel.

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: Epidermal Growth Factor receptor (EGFR) internalises into Glioblastoma multiforme (GBM) cells upon mTOR inhibition. ( A ) U87MG (upper panels), GL15 (middle panels) and primary GH2 cells (lower panels) were cultured in complete medium (DMEM) (CTR) or in DMEM in the presence of 250 nM Torin1 for 18 h. Immunocytochemistry and confocal analysis for EGFR localisation (red) were then performed. Hoechst 33342 was used to stain nuclei (blue). Scale bar, 30 μM. Western blot analysis of P-p70S6K and p70S6K was performed to check mTOR pathway inhibition by Torin1. β-actin was used as loading control. The blots are representative of three independent experiments. ( B ) Immunocytochemistry and confocal analysis for EGFR localisation (red) were performed in U87MG cells, upon 6 h and 24 h Torin1 treatment in the presence or absence of 100 μM Dynasore. Scale bar, 30 μM. A 4× magnification is shown for the right panels representing cells treated with Torin1 plus Dynasore. ( C ) U87MG cells were incubated with 0.5 mg/mL Rhodamine Dextran for the indicated times and its uptake within the cells analysed by image capturing at the confocal microscope. Scale bar, 30 μM. Fluorescence quantification of Dextran uptake at 120′ is shown in the right panel.

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Inhibition, Cell Culture, Immunocytochemistry, Staining, Western Blot, Incubation, Microscopy, Fluorescence

    EGFR de-localisation in GBM cells is independent of canonical autophagy. ( A ) shCTR and shBECLIN1 GL15 cells [ 23 ] were cultured in DMEM (CTR) or amino acid- and serum- free medium (EBSS) media or in DMEM in the presence of 250 nM Torin1 for 18 h and subjected to immunocytochemistry and confocal analysis for EGFR localisation (red) (right panels). Hoechst 33342 was used to stain nuclei (blue). Scale bar, 30 μM. Western blot analysis of P62 and LC3 I/II was also performed in basal conditions to check autophagy status (left panel). A specific antibody for BECLIN1 was used to check the silencing efficiency. β-ACTIN was used as loading control. The blot is representative of three independent experiments. ( B ) U87MG cells were transduced with GFP-LC3-expressing retrovirus as described in Material and Methods. Infected cells, cultured in DMEM alone (CTR) or in DMEM containing 250 nM Torin1 for 24 h, were subjected to immunocytochemistry and confocal analysis for EGFR (red) and autophagosomes (green) localisation. Colocalisation was excluded by calculating the Pearson’s correlation coefficient r (mean r CTR, 0.15 ± 0.02; Torin1, 0.2 ± 0.03). The images showing the merge of the two signals are shown in the right panels. Scale bar, 30 μM.

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: EGFR de-localisation in GBM cells is independent of canonical autophagy. ( A ) shCTR and shBECLIN1 GL15 cells [ 23 ] were cultured in DMEM (CTR) or amino acid- and serum- free medium (EBSS) media or in DMEM in the presence of 250 nM Torin1 for 18 h and subjected to immunocytochemistry and confocal analysis for EGFR localisation (red) (right panels). Hoechst 33342 was used to stain nuclei (blue). Scale bar, 30 μM. Western blot analysis of P62 and LC3 I/II was also performed in basal conditions to check autophagy status (left panel). A specific antibody for BECLIN1 was used to check the silencing efficiency. β-ACTIN was used as loading control. The blot is representative of three independent experiments. ( B ) U87MG cells were transduced with GFP-LC3-expressing retrovirus as described in Material and Methods. Infected cells, cultured in DMEM alone (CTR) or in DMEM containing 250 nM Torin1 for 24 h, were subjected to immunocytochemistry and confocal analysis for EGFR (red) and autophagosomes (green) localisation. Colocalisation was excluded by calculating the Pearson’s correlation coefficient r (mean r CTR, 0.15 ± 0.02; Torin1, 0.2 ± 0.03). The images showing the merge of the two signals are shown in the right panels. Scale bar, 30 μM.

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Cell Culture, Immunocytochemistry, Staining, Western Blot, Transduction, Expressing, Infection

    mTOR inhibition leads to cell proliferation arrest. ( A ) U87MG, GL15 and GH2 cells were cultured in complete DMEM (CTR) or in the presence of 250 nM Torin1 (TORIN1) or 500 µM Temozolomide (TMZ) or both. At the indicated time points, cells were trypsinised and counted in a Newbauer chamber. The graph represents the mean ± Standard Error of the Mean (SEM) of three different experiments. Statistical significance: * indicates significance vs. Ctr; # indicates significance vs. Torin1 + TMZ; ° indicates significance vs. Torin1. ** p

    Journal: Cancers

    Article Title: mTOR Inhibition Leads to Src-Mediated EGFR Internalisation and Degradation in Glioma Cells

    doi: 10.3390/cancers12082266

    Figure Lengend Snippet: mTOR inhibition leads to cell proliferation arrest. ( A ) U87MG, GL15 and GH2 cells were cultured in complete DMEM (CTR) or in the presence of 250 nM Torin1 (TORIN1) or 500 µM Temozolomide (TMZ) or both. At the indicated time points, cells were trypsinised and counted in a Newbauer chamber. The graph represents the mean ± Standard Error of the Mean (SEM) of three different experiments. Statistical significance: * indicates significance vs. Ctr; # indicates significance vs. Torin1 + TMZ; ° indicates significance vs. Torin1. ** p

    Article Snippet: Human U87MG, GL15 were cultured in DMEM (Lonza, Basel, Switzerland), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Euroclone, Milan, Italy) and 1% penicillin/streptomycin solution (Euroclone, Milan, Italy).

    Techniques: Inhibition, Cell Culture