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Cambrex fetal bovine serum fbs
Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in <t>DMEM/F12</t> supplemented with 10% <t>FBS.</t> One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P
Fetal Bovine Serum Fbs, supplied by Cambrex, used in various techniques. Bioz Stars score: 93/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells"

Article Title: Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells

Journal: Nutrition Research and Practice

doi: 10.4162/nrp.2015.9.2.111

Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P
Figure Legend Snippet: Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P

Techniques Used: Incubation, MTT Assay

2) Product Images from "Cyclic AMP Controls mTOR through Regulation of the Dynamic Interaction between Rheb and Phosphodiesterase 4D ▿"

Article Title: Cyclic AMP Controls mTOR through Regulation of the Dynamic Interaction between Rheb and Phosphodiesterase 4D ▿

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.00217-10

cAMP activates mTORC1 signaling via Rheb. (A) cAMP activates mTORC1 signaling in a rapamycin-dependent manner. HEK293 cells were incubated with 10% fetal bovine serum for 36 h. After 24 h of serum deprivation, the cells were preincubated with or without serum-free medium containing 10 nM rapamycin for 45 min and were then treated with 10 μM forskolin for 5 min. The cells were lysed with buffer containing 0.5% CHAPS. Equal amounts of total cell lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. (B) Forskolin activates mTORC1 in a dose-dependent manner. After 24 h of serum deprivation, HEK293 cells were treated with the indicated concentrations of forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. (C) cAMP-mediated mTORC1 activation is dependent on Rheb. HEK293 cells were transfected with either the control or Rheb shRNA by using Lipofectamine. After 24 h, cells were depleted of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were immunoprecipitated (IP) with anti-Rheb antibodies. The immunoprecipitates and lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. CTL, cytotoxic T lymphocytes. (D) The level of cAMP-mediated mTORC1 activation is decreased by a Rheb dominant negative mutant. HEK293 cells were transfected with either a GFP vector or GFP-Rheb D60I by using Lipofectamine. After 24 h, cells were depleted of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (E) cAMP-mediated mTORC1 activation is not affected in TSC1-deficient cells. TSC1 +/+ and TSC1 −/− MEFs were depleted of serum for 8 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (F) Overexpression of TSC1 and TSC2 has little effect on forskolin-dependent mTORC1 activation. HEK293 cells were transfected with control or recombinant TSC1/2. After 24 h, cells were deprived of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (G) Overexpression of the AMPKα1 dominant negative form does not affect forskolin-dependent mTORC1 activation. HEK293 cells were transfected with the control or recombinant form of dominant-negative AMPKα1. After 24 h, cells were deprived of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. DN, dominant negative. (H) Inhibition of MAPK does not affect forskolin-dependent mTORC1 activation. After 24 h of serum deprivation, HEK293 cells were preincubated with or without serum-free medium containing PD98059 for 30 min and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. (I) Inhibition of PKA does not affect forskolin-dependent mTORC1 activation. After 24 h of serum deprivation, HEK293 cells were preincubated with or without serum-free medium containing H89 for 45 min and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments.
Figure Legend Snippet: cAMP activates mTORC1 signaling via Rheb. (A) cAMP activates mTORC1 signaling in a rapamycin-dependent manner. HEK293 cells were incubated with 10% fetal bovine serum for 36 h. After 24 h of serum deprivation, the cells were preincubated with or without serum-free medium containing 10 nM rapamycin for 45 min and were then treated with 10 μM forskolin for 5 min. The cells were lysed with buffer containing 0.5% CHAPS. Equal amounts of total cell lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. (B) Forskolin activates mTORC1 in a dose-dependent manner. After 24 h of serum deprivation, HEK293 cells were treated with the indicated concentrations of forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. (C) cAMP-mediated mTORC1 activation is dependent on Rheb. HEK293 cells were transfected with either the control or Rheb shRNA by using Lipofectamine. After 24 h, cells were depleted of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were immunoprecipitated (IP) with anti-Rheb antibodies. The immunoprecipitates and lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. CTL, cytotoxic T lymphocytes. (D) The level of cAMP-mediated mTORC1 activation is decreased by a Rheb dominant negative mutant. HEK293 cells were transfected with either a GFP vector or GFP-Rheb D60I by using Lipofectamine. After 24 h, cells were depleted of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (E) cAMP-mediated mTORC1 activation is not affected in TSC1-deficient cells. TSC1 +/+ and TSC1 −/− MEFs were depleted of serum for 8 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (F) Overexpression of TSC1 and TSC2 has little effect on forskolin-dependent mTORC1 activation. HEK293 cells were transfected with control or recombinant TSC1/2. After 24 h, cells were deprived of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of three independent experiments. (G) Overexpression of the AMPKα1 dominant negative form does not affect forskolin-dependent mTORC1 activation. HEK293 cells were transfected with the control or recombinant form of dominant-negative AMPKα1. After 24 h, cells were deprived of serum for 24 h and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. DN, dominant negative. (H) Inhibition of MAPK does not affect forskolin-dependent mTORC1 activation. After 24 h of serum deprivation, HEK293 cells were preincubated with or without serum-free medium containing PD98059 for 30 min and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments. (I) Inhibition of PKA does not affect forskolin-dependent mTORC1 activation. After 24 h of serum deprivation, HEK293 cells were preincubated with or without serum-free medium containing H89 for 45 min and then treated with 10 μM forskolin for 5 min. The lysates were subjected to SDS-PAGE and immunoblotted with the respective antibodies. The results shown are representative of two independent experiments.

Techniques Used: Incubation, SDS Page, Activation Assay, Transfection, shRNA, Immunoprecipitation, CTL Assay, Dominant Negative Mutation, Plasmid Preparation, Over Expression, Recombinant, Inhibition

3) Product Images from "Caveolar Endocytosis Is Critical for BK Virus Infection of Human Renal Proximal Tubular Epithelial Cells ▿"

Article Title: Caveolar Endocytosis Is Critical for BK Virus Infection of Human Renal Proximal Tubular Epithelial Cells ▿

Journal:

doi: 10.1128/JVI.00924-07

MBCD interfered with BKV infection. HRPTEC were preincubated with MBCD (1.25, 2.5, and 5 mM) for 1 h prior to coincubation with BKV (MOI, 0.5 FFU/cell). After 72 h, medium was removed and cells were washed three times with REBM with 0.5% FBS and
Figure Legend Snippet: MBCD interfered with BKV infection. HRPTEC were preincubated with MBCD (1.25, 2.5, and 5 mM) for 1 h prior to coincubation with BKV (MOI, 0.5 FFU/cell). After 72 h, medium was removed and cells were washed three times with REBM with 0.5% FBS and

Techniques Used: Infection

4) Product Images from "Differences in Cytotoxic, Genotoxic, and Inflammatory Response of Bronchial and Alveolar Human Lung Epithelial Cells to Pristine and COOH-Functionalized Multiwalled Carbon Nanotubes"

Article Title: Differences in Cytotoxic, Genotoxic, and Inflammatory Response of Bronchial and Alveolar Human Lung Epithelial Cells to Pristine and COOH-Functionalized Multiwalled Carbon Nanotubes

Journal: BioMed Research International

doi: 10.1155/2014/359506

Percent association of pristine and COOH-functionalized MWCNTs with A549 cells in RPMI cell medium with 10% FBS (a) and with BEAS-2B cells in BEGM cell medium (b). The value of cellular uptake was expressed as percentage of total MWCNT added to the well (mean ± SD). ∗ P ≤ 0.05, MWCNT-COOH versus MWCNTs.
Figure Legend Snippet: Percent association of pristine and COOH-functionalized MWCNTs with A549 cells in RPMI cell medium with 10% FBS (a) and with BEAS-2B cells in BEGM cell medium (b). The value of cellular uptake was expressed as percentage of total MWCNT added to the well (mean ± SD). ∗ P ≤ 0.05, MWCNT-COOH versus MWCNTs.

Techniques Used:

Representative optical microscopy images of A549 cells exposed for 24 h to CNTs before washing them to remove CNTs dissolved in the medium. (a) Cells exposed to 40 μ g/mL of pristine MWCNTs; (b) cells exposed to 40 μ g/mL of MWCNT-COOH; (c) control cells. Magnification: 20x. TEM micrographs of pristine MWCNTs (d) and MWCNT-COOH (e) agglomerates/aggregates in RPMI with 10% FBS culture medium (bar 2 μ m).
Figure Legend Snippet: Representative optical microscopy images of A549 cells exposed for 24 h to CNTs before washing them to remove CNTs dissolved in the medium. (a) Cells exposed to 40 μ g/mL of pristine MWCNTs; (b) cells exposed to 40 μ g/mL of MWCNT-COOH; (c) control cells. Magnification: 20x. TEM micrographs of pristine MWCNTs (d) and MWCNT-COOH (e) agglomerates/aggregates in RPMI with 10% FBS culture medium (bar 2 μ m).

Techniques Used: Microscopy, Transmission Electron Microscopy

5) Product Images from "Chestnut extract induces apoptosis in AGS human gastric cancer cells"

Article Title: Chestnut extract induces apoptosis in AGS human gastric cancer cells

Journal: Nutrition Research and Practice

doi: 10.4162/nrp.2011.5.3.185

Effect of an ethanol extract of chestnut powder (CPE) on AGS cell viability and apoptosis. AGS cells were plated at a density of 50,000 cells/well in 24-well plates with DMEM/F12 supplemented with 10% FBS. One day after plating, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% FBS serum-deprivation medium for 24 h. After serum deprivation, cells were incubated in serum-deprivation medium in the absence or presence of various concentrations of CPE. (A) Cell numbers were estimated by the MTT assay. (B) Apoptotic cells were detected with the Cell death detection ELISA PLUS . Each bar represents the mean ± SEM (n = 6). Means without a common letter differ significantly, P
Figure Legend Snippet: Effect of an ethanol extract of chestnut powder (CPE) on AGS cell viability and apoptosis. AGS cells were plated at a density of 50,000 cells/well in 24-well plates with DMEM/F12 supplemented with 10% FBS. One day after plating, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% FBS serum-deprivation medium for 24 h. After serum deprivation, cells were incubated in serum-deprivation medium in the absence or presence of various concentrations of CPE. (A) Cell numbers were estimated by the MTT assay. (B) Apoptotic cells were detected with the Cell death detection ELISA PLUS . Each bar represents the mean ± SEM (n = 6). Means without a common letter differ significantly, P

Techniques Used: Incubation, MTT Assay, Enzyme-linked Immunosorbent Assay

Effect of chestnut extracts on the viable cell numbers of MDA-MD-231, DU145, and AGS cells. (A) MDA-MB-231, (B) DU145, and (C) AGS cells were plated at a density of 50,000 cells/well in 24-well plates with DMEM/F12 supplemented with 10% FBS. One day after plating, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% FBS serum-deprivation medium for 24 h. After serum deprivation, the cells were incubated in serum-deprivation medium in the absence or presence of 200 µg/mL viscous, aqueous extract of raw chestnut (RCW1), non-viscous, aqueous extract of raw chestnut (RCW2), aqueous extract of chestnut powder (CPW), ethanol extract of raw chestnut (RCE), or an ethanol extract of chestnut powder (CPE) for 24 hr. Cell numbers were estimated by the MTT assay. Each bar represents the mean ± SEM (n = 6). Means at a time without a common letter differ significantly, P
Figure Legend Snippet: Effect of chestnut extracts on the viable cell numbers of MDA-MD-231, DU145, and AGS cells. (A) MDA-MB-231, (B) DU145, and (C) AGS cells were plated at a density of 50,000 cells/well in 24-well plates with DMEM/F12 supplemented with 10% FBS. One day after plating, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% FBS serum-deprivation medium for 24 h. After serum deprivation, the cells were incubated in serum-deprivation medium in the absence or presence of 200 µg/mL viscous, aqueous extract of raw chestnut (RCW1), non-viscous, aqueous extract of raw chestnut (RCW2), aqueous extract of chestnut powder (CPW), ethanol extract of raw chestnut (RCE), or an ethanol extract of chestnut powder (CPE) for 24 hr. Cell numbers were estimated by the MTT assay. Each bar represents the mean ± SEM (n = 6). Means at a time without a common letter differ significantly, P

Techniques Used: Multiple Displacement Amplification, Incubation, MTT Assay

6) Product Images from "Fucoidan present in brown algae induces apoptosis of human colon cancer cells"

Article Title: Fucoidan present in brown algae induces apoptosis of human colon cancer cells

Journal: BMC Gastroenterology

doi: 10.1186/1471-230X-10-96

Fucoidan reduces the viability of HT-29 and HCT116 cells . HT-29 (A) , HCT116 (B) and FHC (C) cells were plated in 24-well plates at a density of 50,000 cells/well with DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% charcoal-stripped FBS (serum-deprivation medium) for 24 h. Following serum deprivation, the cells were incubated in serum-deprivation medium containing 0 - 20 μg/mL of fucoidan. Viable cell numbers were estimated via MTT assays. Each bar represents the mean ± SEM (n = 6). Means at a time without a common letter differ, P
Figure Legend Snippet: Fucoidan reduces the viability of HT-29 and HCT116 cells . HT-29 (A) , HCT116 (B) and FHC (C) cells were plated in 24-well plates at a density of 50,000 cells/well with DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-deprived with DMEM/F12 supplemented with 1% charcoal-stripped FBS (serum-deprivation medium) for 24 h. Following serum deprivation, the cells were incubated in serum-deprivation medium containing 0 - 20 μg/mL of fucoidan. Viable cell numbers were estimated via MTT assays. Each bar represents the mean ± SEM (n = 6). Means at a time without a common letter differ, P

Techniques Used: Incubation, MTT Assay

Related Articles

Modification:

Article Title: Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells
Article Snippet: .. Materials The reagents used in this study were purchased from the following suppliers: Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture (DMEM/F12) and selenium from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS), trypsin-EDTA, and penicillin/streptomycin from Cambrex Bio Technology (Walkersville, MD, USA); 3-[4,5-dimetjylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT), anti-β-actin, RIA-grade bovine serum albumin (BSA), and transferrin from Sigma-Aldrich Co. (St. Louis, MO, USA); antibodies against cyclin D1 and phospho-Rb (Ser807/811) from Cell Signaling Technology (Beverly, MA, USA); antibodies against p21CIP1/WAF1 (c-19), p27KIP1 , p53, CDK2 (M-2), CDK4 (c-22), E2F-1 (C-20), and Rb (c-15) from Santa Cruz Biotechnology (Santa Cruz, CA, USA). ..

Article Title: Chestnut extract induces apoptosis in AGS human gastric cancer cells
Article Snippet: .. The reagents employed in this study were purchased from the indicated suppliers: Anti-β-actin antibody and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA); Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient mixture (DMEM/F-12) was purchased from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS) was obtained from Cambrex Bio Technology (Walkersville, MD, USA); horse-radish peroxidase (HRP)-conjugated anti-rabbit and anti-mouse IgG were purchased from Amersham Biosciences (Arlington Heights, IL, USA); antibodies against cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, cleaved poly (ADP-ribose) polymerase (PARP), caspase-8, Bid, and X-linked inhibitor of apoptosis protein (XIAP) were obtained from Cell Signaling Technology (Beverly, MA, USA); antibody against tumor necrosis factor-related apoptosis including ligand (TRAIL) was obtained from BD Pharmingen (Franklin Lake, NJ, USA); antibodies against Fas, Fas ligand (Fas-L), Bcl-2, and Bax were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and antibodies against death receptor (DR) 4 and 5 were obtained from Imgenex (San Diego, CA, USA). ..

Article Title: Molecular cloning and anti-invasive activity of cathepsin L propeptide-like protein from Calotropis procera R. Br. against cancer cells
Article Snippet: .. Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were purchased from Cambrex (Walkersville, MD). .. Cysteine protease unigenes from Calotropis procera R. Br. were translated over six frames by the ExPASy translate tool and protein functional domains were predicted by InterProScan 4 web program .

Article Title: Fucoidan present in brown algae induces apoptosis of human colon cancer cells
Article Snippet: .. Materials The reagents employed in this study were purchased from the indicated suppliers: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), biobenzimide H 33258 (Hoechst 33258), Z-IETD-FMK, Z-LEHD-FMK, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1), anti-β-actin antibody, and anti-α-tubulin antibody (Sigma-Aldrich Co.); Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient mixture (DMEM/F-12) (Gibco BRL, Gaithersburg, MD, USA); fetal bovine serum (FBS) (Cambrex Bio Technology, Walkersville, MD, USA); a horseradish peroxidase (HRP)-conjugated anti-rabbit, anti-goat, and anti-mouse IgG (Amersham Biosciences, Arlington Heights, IL, USA); antibodies against cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, cleaved poly (ADP-ribose) polymerase (PARP), caspase-8, Bid, survivin, and X-linked inhibitor of apoptosis protein (XIAP) (Cell Signaling Technology, Beverly, MA, USA); phycoerythrin-conjugated Annexin V (PE-Annexin V), 7-amino-actinomycin D (7-AAD), and antibodies against cytochrome c and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) (BD Pharmingen, Franklin Lake, NJ, USA); antibodies against Bcl-2, Bax, Fas, Fas ligand (FasL), Smac/Diablo, and heat shock protein (HSP) 60 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); antibodies against death receptor (DR) 4 and 5 (Imgenex, San Diego, CA, USA). ..

MTT Assay:

Article Title: Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells
Article Snippet: .. Materials The reagents used in this study were purchased from the following suppliers: Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture (DMEM/F12) and selenium from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS), trypsin-EDTA, and penicillin/streptomycin from Cambrex Bio Technology (Walkersville, MD, USA); 3-[4,5-dimetjylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT), anti-β-actin, RIA-grade bovine serum albumin (BSA), and transferrin from Sigma-Aldrich Co. (St. Louis, MO, USA); antibodies against cyclin D1 and phospho-Rb (Ser807/811) from Cell Signaling Technology (Beverly, MA, USA); antibodies against p21CIP1/WAF1 (c-19), p27KIP1 , p53, CDK2 (M-2), CDK4 (c-22), E2F-1 (C-20), and Rb (c-15) from Santa Cruz Biotechnology (Santa Cruz, CA, USA). ..

Article Title: Chestnut extract induces apoptosis in AGS human gastric cancer cells
Article Snippet: .. The reagents employed in this study were purchased from the indicated suppliers: Anti-β-actin antibody and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA); Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient mixture (DMEM/F-12) was purchased from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS) was obtained from Cambrex Bio Technology (Walkersville, MD, USA); horse-radish peroxidase (HRP)-conjugated anti-rabbit and anti-mouse IgG were purchased from Amersham Biosciences (Arlington Heights, IL, USA); antibodies against cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, cleaved poly (ADP-ribose) polymerase (PARP), caspase-8, Bid, and X-linked inhibitor of apoptosis protein (XIAP) were obtained from Cell Signaling Technology (Beverly, MA, USA); antibody against tumor necrosis factor-related apoptosis including ligand (TRAIL) was obtained from BD Pharmingen (Franklin Lake, NJ, USA); antibodies against Fas, Fas ligand (Fas-L), Bcl-2, and Bax were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and antibodies against death receptor (DR) 4 and 5 were obtained from Imgenex (San Diego, CA, USA). ..

Article Title: Fucoidan present in brown algae induces apoptosis of human colon cancer cells
Article Snippet: .. Materials The reagents employed in this study were purchased from the indicated suppliers: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), biobenzimide H 33258 (Hoechst 33258), Z-IETD-FMK, Z-LEHD-FMK, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1), anti-β-actin antibody, and anti-α-tubulin antibody (Sigma-Aldrich Co.); Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient mixture (DMEM/F-12) (Gibco BRL, Gaithersburg, MD, USA); fetal bovine serum (FBS) (Cambrex Bio Technology, Walkersville, MD, USA); a horseradish peroxidase (HRP)-conjugated anti-rabbit, anti-goat, and anti-mouse IgG (Amersham Biosciences, Arlington Heights, IL, USA); antibodies against cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, cleaved poly (ADP-ribose) polymerase (PARP), caspase-8, Bid, survivin, and X-linked inhibitor of apoptosis protein (XIAP) (Cell Signaling Technology, Beverly, MA, USA); phycoerythrin-conjugated Annexin V (PE-Annexin V), 7-amino-actinomycin D (7-AAD), and antibodies against cytochrome c and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) (BD Pharmingen, Franklin Lake, NJ, USA); antibodies against Bcl-2, Bax, Fas, Fas ligand (FasL), Smac/Diablo, and heat shock protein (HSP) 60 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); antibodies against death receptor (DR) 4 and 5 (Imgenex, San Diego, CA, USA). ..

Cell Culture:

Article Title: Differences in Cytotoxic, Genotoxic, and Inflammatory Response of Bronchial and Alveolar Human Lung Epithelial Cells to Pristine and COOH-Functionalized Multiwalled Carbon Nanotubes
Article Snippet: .. A549 cells were cultured in Rosewell Park Memorial Institute 1640 medium (RPMI 1640) (EuroClone, United Kingdom) supplemented with 10% fetal bovine serum (FBS) and BEAS-2B cells were cultured in Bronchial Epithelial cell Growth Medium (BEGM) BulletKit (Cambrex Bio Science Walkersville Inc.) both at 37°C in 5% CO2 . .. Cells (8 × 104 cells/well) were seeded into 24-multiwell culture plate (15.6 mm well diameter) and cultured for 24 h before the exposure.

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    Cambrex xvivo 10
    Morphology of monocyte-derived macrophages after 6 days. Differentiation in the presence of GM-CSF ( A ) or M-CSF ( B ) in <t>Xvivo</t> 10 or RPMI +10% FBS. ( C ) Differentiation in XVivo 10 media supplemented with the indicated cytokines, as described in Material and Methods . The scale denotes 50 µm.
    Xvivo 10, supplied by Cambrex, used in various techniques. Bioz Stars score: 89/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cambrex heat inactivated low endotoxin fetal bovine serum fbs
    (1–4) Dose-dependent response for anti-proliferative properties of various compounds in cancer cells of Hela-1, liver-2, pancreas-3, and prostate-4. The cancer cell lines of Hela, liver, pancreas, and prostate were maintained in <t>DMEM</t> supplemented with 10% heat inactivated <t>FBS</t> and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. Cancer cells (1 × 10 5 ) of various organs were seeded in 48 well tissue culture plate with 900 μl of medium containing 0.2% dimethyl sulfoxide of different types of cancer cell lines (Hela cell, liver, pancreas, and prostate), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate were added to each well, incubated for 48 h at 37 °C in a humidified atmosphere of 5% CO 2 . Followed by counting of live cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT), as described previously [ 18 , 19 ]. The anticancer properties and dose-dependence for eleven compounds are presented for Hela, liver, pancreas, and prostate cancer cell lines. Values in a column not sharing a common symbol are significantly different at ¶ = P
    Heat Inactivated Low Endotoxin Fetal Bovine Serum Fbs, supplied by Cambrex, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 5 article reviews
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    Cambrex fetal bovine serum fbs
    Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in <t>DMEM/F12</t> supplemented with 10% <t>FBS.</t> One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P
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    Morphology of monocyte-derived macrophages after 6 days. Differentiation in the presence of GM-CSF ( A ) or M-CSF ( B ) in Xvivo 10 or RPMI +10% FBS. ( C ) Differentiation in XVivo 10 media supplemented with the indicated cytokines, as described in Material and Methods . The scale denotes 50 µm.

    Journal: PLoS ONE

    Article Title: In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

    doi: 10.1371/journal.pone.0042656

    Figure Lengend Snippet: Morphology of monocyte-derived macrophages after 6 days. Differentiation in the presence of GM-CSF ( A ) or M-CSF ( B ) in Xvivo 10 or RPMI +10% FBS. ( C ) Differentiation in XVivo 10 media supplemented with the indicated cytokines, as described in Material and Methods . The scale denotes 50 µm.

    Article Snippet: Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act.

    Techniques: Derivative Assay

    Cytokine concentration in supernatant of monocyte-derived macrophages. Monocytes were stimulated for 6 days with ( A ) GM-CSF (M1) or M-CSF (M2) in XVivo 10 or RPMI +10% FBS, or with ( B ) GM-CSF for 3 days and LPS and IFN-γ for 3 additional days (M1), M-CSF+IL-4 (M2a) or M-CSF+IL-10 (M2c) in XVivo 10. Data represent mean ± SEM of cytokine concentration of at least 8 donors. Statistical significance was determined using Tukey-Kramer HSD test pairwise comparison (** p

    Journal: PLoS ONE

    Article Title: In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

    doi: 10.1371/journal.pone.0042656

    Figure Lengend Snippet: Cytokine concentration in supernatant of monocyte-derived macrophages. Monocytes were stimulated for 6 days with ( A ) GM-CSF (M1) or M-CSF (M2) in XVivo 10 or RPMI +10% FBS, or with ( B ) GM-CSF for 3 days and LPS and IFN-γ for 3 additional days (M1), M-CSF+IL-4 (M2a) or M-CSF+IL-10 (M2c) in XVivo 10. Data represent mean ± SEM of cytokine concentration of at least 8 donors. Statistical significance was determined using Tukey-Kramer HSD test pairwise comparison (** p

    Article Snippet: Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act.

    Techniques: Concentration Assay, Derivative Assay

    Receptor expression level on monocyte-derived macrophages. Surface expression of CD163 (A) or CD206 (B) on M1 and M2 MDM in Xvivo 10 or RPMI +10% FBS, or on M1, M2a and M2c MDM in XVivo 10 media. Data represent mean ± SEM of Mean Fluorescence Intensity (Geom. mean) of at least 8 donors. Statistical significance was determined using Tukey-Kramer HSD test pairwise comparison (***p

    Journal: PLoS ONE

    Article Title: In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

    doi: 10.1371/journal.pone.0042656

    Figure Lengend Snippet: Receptor expression level on monocyte-derived macrophages. Surface expression of CD163 (A) or CD206 (B) on M1 and M2 MDM in Xvivo 10 or RPMI +10% FBS, or on M1, M2a and M2c MDM in XVivo 10 media. Data represent mean ± SEM of Mean Fluorescence Intensity (Geom. mean) of at least 8 donors. Statistical significance was determined using Tukey-Kramer HSD test pairwise comparison (***p

    Article Snippet: Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act.

    Techniques: Expressing, Derivative Assay, Fluorescence

    Phagocytic activity of M1, M2a and M2c monocyte-derived macrophages towards MOLT4. Gating strategy used to assess phagocytosis of MOLT4 debris by MDM obtained by flow cytometry: controls included assessment of MOLT4 alone or together with MDM in PBS ( A ). Dot plots representing data from two distinct donors in XVivo 10 as described in Methods ( B ). Mean fluorescence intensity (median) of phagocytosing MDM derived from ten distinct donors, attributed with identical (□) or differential (• or ○) activity between activated M1 and M2a or M2c MDM ( C ). Horizontal bars represent the median value of each group.

    Journal: PLoS ONE

    Article Title: In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

    doi: 10.1371/journal.pone.0042656

    Figure Lengend Snippet: Phagocytic activity of M1, M2a and M2c monocyte-derived macrophages towards MOLT4. Gating strategy used to assess phagocytosis of MOLT4 debris by MDM obtained by flow cytometry: controls included assessment of MOLT4 alone or together with MDM in PBS ( A ). Dot plots representing data from two distinct donors in XVivo 10 as described in Methods ( B ). Mean fluorescence intensity (median) of phagocytosing MDM derived from ten distinct donors, attributed with identical (□) or differential (• or ○) activity between activated M1 and M2a or M2c MDM ( C ). Horizontal bars represent the median value of each group.

    Article Snippet: Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act.

    Techniques: Activity Assay, Derivative Assay, Flow Cytometry, Cytometry, Fluorescence

    Effect of monocyte-derived macrophage supernatant on HCC1143 cell line proliferation. Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act. M1), M2a and M2c MDM stimulated in XVivo 10. Identical y-axis scales were used for comparison sake. Data represent mean ± SEM of three independent experiments including each condition in triplicate. Statistical significance was determined using t-test pairwise comparison (** p

    Journal: PLoS ONE

    Article Title: In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

    doi: 10.1371/journal.pone.0042656

    Figure Lengend Snippet: Effect of monocyte-derived macrophage supernatant on HCC1143 cell line proliferation. Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act. M1), M2a and M2c MDM stimulated in XVivo 10. Identical y-axis scales were used for comparison sake. Data represent mean ± SEM of three independent experiments including each condition in triplicate. Statistical significance was determined using t-test pairwise comparison (** p

    Article Snippet: Evaluation was performed in the presence or the absence of neutralizing TNF-α antibodies, in ( A ) conditioned media from M1 or M2 MDM cultured in RPMI 10% FBS or XVivo 10, ( B ) conditioned media from activated M1 (act.

    Techniques: Derivative Assay, Cell Culture, Activated Clotting Time Assay

    (1–4) Dose-dependent response for anti-proliferative properties of various compounds in cancer cells of Hela-1, liver-2, pancreas-3, and prostate-4. The cancer cell lines of Hela, liver, pancreas, and prostate were maintained in DMEM supplemented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. Cancer cells (1 × 10 5 ) of various organs were seeded in 48 well tissue culture plate with 900 μl of medium containing 0.2% dimethyl sulfoxide of different types of cancer cell lines (Hela cell, liver, pancreas, and prostate), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate were added to each well, incubated for 48 h at 37 °C in a humidified atmosphere of 5% CO 2 . Followed by counting of live cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT), as described previously [ 18 , 19 ]. The anticancer properties and dose-dependence for eleven compounds are presented for Hela, liver, pancreas, and prostate cancer cell lines. Values in a column not sharing a common symbol are significantly different at ¶ = P

    Journal: Lipids in Health and Disease

    Article Title: Proteasome inhibitors modulate anticancer and anti-proliferative properties via NF-kB signaling, and ubiquitin-proteasome pathways in cancer cell lines of different organs

    doi: 10.1186/s12944-018-0697-5

    Figure Lengend Snippet: (1–4) Dose-dependent response for anti-proliferative properties of various compounds in cancer cells of Hela-1, liver-2, pancreas-3, and prostate-4. The cancer cell lines of Hela, liver, pancreas, and prostate were maintained in DMEM supplemented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. Cancer cells (1 × 10 5 ) of various organs were seeded in 48 well tissue culture plate with 900 μl of medium containing 0.2% dimethyl sulfoxide of different types of cancer cell lines (Hela cell, liver, pancreas, and prostate), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate were added to each well, incubated for 48 h at 37 °C in a humidified atmosphere of 5% CO 2 . Followed by counting of live cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT), as described previously [ 18 , 19 ]. The anticancer properties and dose-dependence for eleven compounds are presented for Hela, liver, pancreas, and prostate cancer cell lines. Values in a column not sharing a common symbol are significantly different at ¶ = P

    Article Snippet: Materials Dulbecco’s Modified Eagle Medium (DMEM), heat-inactivated low-endotoxin fetal bovine serum (FBS), and gentamicin purchased from Cambrex (Walkersville MD, USA) for tissue culture.

    Techniques: Incubation, Colorimetric Assay, MTT Assay

    (9) Dose-dependent response for anti-proliferative properties of various compounds in T-cells (Jurkat). The T-cells (Jurkat) maintained in DMEM supplemented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. The T-cells (1 × 10 5 ) of was seeded in 48 well tissue culture plate with 900 μl of medium, containing 0.2% dimethyl sulfoxide of T-cells (Jurkat), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate added in each well, and then incubated for 48 h at 37 °C, in a humidified atmosphere of 5% CO 2 . Followed by counting the living cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT) as described previously [ 18 , 19 ]. The anti-proliferation properties as dose-dependent with eleven compounds were presented in T-cells (Jurkat). Values in a column not sharing a common symbol are significantly different at ¶ = P

    Journal: Lipids in Health and Disease

    Article Title: Proteasome inhibitors modulate anticancer and anti-proliferative properties via NF-kB signaling, and ubiquitin-proteasome pathways in cancer cell lines of different organs

    doi: 10.1186/s12944-018-0697-5

    Figure Lengend Snippet: (9) Dose-dependent response for anti-proliferative properties of various compounds in T-cells (Jurkat). The T-cells (Jurkat) maintained in DMEM supplemented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. The T-cells (1 × 10 5 ) of was seeded in 48 well tissue culture plate with 900 μl of medium, containing 0.2% dimethyl sulfoxide of T-cells (Jurkat), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate added in each well, and then incubated for 48 h at 37 °C, in a humidified atmosphere of 5% CO 2 . Followed by counting the living cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT) as described previously [ 18 , 19 ]. The anti-proliferation properties as dose-dependent with eleven compounds were presented in T-cells (Jurkat). Values in a column not sharing a common symbol are significantly different at ¶ = P

    Article Snippet: Materials Dulbecco’s Modified Eagle Medium (DMEM), heat-inactivated low-endotoxin fetal bovine serum (FBS), and gentamicin purchased from Cambrex (Walkersville MD, USA) for tissue culture.

    Techniques: Incubation, Colorimetric Assay, MTT Assay

    (5–8) Dose-dependent response for anti-proliferative properties of various compounds in cancer cell lines of breast-5, lung-6, Melanoma-7, and B-lymphocytes-8. The cancer cell lines of breast, lung, melanoma, and B-lymphocytes were maintained in DMEM supplem ented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. Cancer cells (1 × 10 5 ) of various organs were seeded in 48 well tissue culture plate with 900 μl of medium containing 0.2% dimethyl sulfoxide of different type of cancer cell lines (breast, lung, melanoma, and B-lymphocytes), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate added in each well, and then incubated for 48 h at 37 °C in a humidified atmosphere of 5% CO 2 . Followed by counting the living cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT) as described previously [ 18 , 19 ]. The anticancer properties as dose-dependent of eleven compounds presented for breast, lung, melanoma, and B-lymphocytes cancer cell lines. Values in a column not sharing a common symbol are significantly different at ¶ = P

    Journal: Lipids in Health and Disease

    Article Title: Proteasome inhibitors modulate anticancer and anti-proliferative properties via NF-kB signaling, and ubiquitin-proteasome pathways in cancer cell lines of different organs

    doi: 10.1186/s12944-018-0697-5

    Figure Lengend Snippet: (5–8) Dose-dependent response for anti-proliferative properties of various compounds in cancer cell lines of breast-5, lung-6, Melanoma-7, and B-lymphocytes-8. The cancer cell lines of breast, lung, melanoma, and B-lymphocytes were maintained in DMEM supplem ented with 10% heat inactivated FBS and 10 mg/mL, gentamicin at 37 °C in a humidified atmosphere with 5% carbon dioxide (CO 2 ) and 95% oxygen (O 2 ) as described previously [ 17 ]. Cancer cells (1 × 10 5 ) of various organs were seeded in 48 well tissue culture plate with 900 μl of medium containing 0.2% dimethyl sulfoxide of different type of cancer cell lines (breast, lung, melanoma, and B-lymphocytes), and incubated at 37 °C for 2 h. After 2 h, different concentrations (100 μl of 2.5, 5, 10, 20, 40, or 80 μM) of thiostrepton, ampicillin, dexamethasone, 2-methoxyestradiol, δ-tocotrienol, (−) riboflavin, ascorbic acid, quercetin, amiloride, and quinine sulphate in triplicate added in each well, and then incubated for 48 h at 37 °C in a humidified atmosphere of 5% CO 2 . Followed by counting the living cells of each well by trypan blue dye exclusion or a quantitative colorimetric assay with 3-(4, 5)-dimethylthiozol-2, 5-diphenyl-tetrazolium bromide (MTT) as described previously [ 18 , 19 ]. The anticancer properties as dose-dependent of eleven compounds presented for breast, lung, melanoma, and B-lymphocytes cancer cell lines. Values in a column not sharing a common symbol are significantly different at ¶ = P

    Article Snippet: Materials Dulbecco’s Modified Eagle Medium (DMEM), heat-inactivated low-endotoxin fetal bovine serum (FBS), and gentamicin purchased from Cambrex (Walkersville MD, USA) for tissue culture.

    Techniques: Incubation, Colorimetric Assay, MTT Assay

    Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P

    Journal: Nutrition Research and Practice

    Article Title: Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells

    doi: 10.4162/nrp.2015.9.2.111

    Figure Lengend Snippet: Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P

    Article Snippet: Materials The reagents used in this study were purchased from the following suppliers: Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture (DMEM/F12) and selenium from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS), trypsin-EDTA, and penicillin/streptomycin from Cambrex Bio Technology (Walkersville, MD, USA); 3-[4,5-dimetjylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT), anti-β-actin, RIA-grade bovine serum albumin (BSA), and transferrin from Sigma-Aldrich Co. (St. Louis, MO, USA); antibodies against cyclin D1 and phospho-Rb (Ser807/811) from Cell Signaling Technology (Beverly, MA, USA); antibodies against p21CIP1/WAF1 (c-19), p27KIP1 , p53, CDK2 (M-2), CDK4 (c-22), E2F-1 (C-20), and Rb (c-15) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Incubation, MTT Assay