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  • 99
    Millipore protease inhibitor cocktail
    Protease Inhibitor Cocktail, supplied by Millipore, 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/protease inhibitor cocktail/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail - by Bioz Stars, 2021-04
    99/100 stars
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    86
    Roche protease inhibitor cocktail
    Protease Inhibitor Cocktail, supplied by Roche, 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/protease inhibitor cocktail/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail - by Bioz Stars, 2021-04
    86/100 stars
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    86
    Roche complete protease inhibitor cocktail
    Transport pH profiling of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in <t>complete</t> FF RPMI 1640 medium containing 10% FBS. Twenty-four hours later, DOX was added at 10 ng/ml. After 48 h, transport was measured over 2 min at 37 °C with [ 3 H]MTX (0.5 µM) in MBS (20 mM MES, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 5.5, 6.0 and 6.5) and HBS (20 mM Hepes, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 6.8, 7.0, 7.2 and 7.4). The dishes were washed (3×) with ice-cold PBS. The cells were solubilized in 0.5 N NaOH, and the radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity is calculated in units of pmol [ 3 H]MTX per mg of cell protein and results are presented as mean values plus/minus SDs (Panel A, B) or relative values (relative to the maximum activity for each transporter) as mean plus/minus SDs (Panel C, D) from at least 3 experiments. ( A ) Net transport activities of R1-11/Tet-on-RFC/PCFT cells (in the presence of DOX) are shown over a range of pHs. ( B ) The results depict transport activities of R1-11/Tet-on-RFC/PCFT cells at pH 7.2 (optimum for RFC transport) in the absence or presence of 10 μM PT523 (left panel), and at pH 5.5 (optimum for PCFT transport) in the absence or presence of 10 μM AGF94 (right panel). ( C ) RFC transport activities of R1-11/Tet-on-RFC cells (with DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of PCFT specific inhibitor AGF94 of 10 μM; right panel) are shown over a range of pHs. ( D ) PCFT transport activities of R1-11/Tet-on-RFC/PCFT cells (without DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of RFC specific inhibitor PT523 at 10 μM; right panel) are shown over a range of pHs.
    Complete Protease Inhibitor Cocktail, supplied by Roche, 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/complete protease inhibitor cocktail/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    complete protease inhibitor cocktail - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    99
    Millipore protease inhibitors
    Transport pH profiling of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in <t>complete</t> FF RPMI 1640 medium containing 10% FBS. Twenty-four hours later, DOX was added at 10 ng/ml. After 48 h, transport was measured over 2 min at 37 °C with [ 3 H]MTX (0.5 µM) in MBS (20 mM MES, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 5.5, 6.0 and 6.5) and HBS (20 mM Hepes, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 6.8, 7.0, 7.2 and 7.4). The dishes were washed (3×) with ice-cold PBS. The cells were solubilized in 0.5 N NaOH, and the radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity is calculated in units of pmol [ 3 H]MTX per mg of cell protein and results are presented as mean values plus/minus SDs (Panel A, B) or relative values (relative to the maximum activity for each transporter) as mean plus/minus SDs (Panel C, D) from at least 3 experiments. ( A ) Net transport activities of R1-11/Tet-on-RFC/PCFT cells (in the presence of DOX) are shown over a range of pHs. ( B ) The results depict transport activities of R1-11/Tet-on-RFC/PCFT cells at pH 7.2 (optimum for RFC transport) in the absence or presence of 10 μM PT523 (left panel), and at pH 5.5 (optimum for PCFT transport) in the absence or presence of 10 μM AGF94 (right panel). ( C ) RFC transport activities of R1-11/Tet-on-RFC cells (with DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of PCFT specific inhibitor AGF94 of 10 μM; right panel) are shown over a range of pHs. ( D ) PCFT transport activities of R1-11/Tet-on-RFC/PCFT cells (without DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of RFC specific inhibitor PT523 at 10 μM; right panel) are shown over a range of pHs.
    Protease Inhibitors, supplied by Millipore, 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/protease inhibitors/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protease inhibitors - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier


    N/A
    A cocktail of three protease inhibitors with specificity for the inhibition of metalloproteases cysteine proteases including cathepsins and papain This cocktail is recommended for inhibition of calcium dependent endopeptidases trypsin
      Buy from Supplier

    N/A
    A cocktail of six protease inhibitors with broad specificity for the inhibition of aspartic serine cysteine aminopeptidases and metalloproteases This cocktail is recommended for use with plant cell extracts Each
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    Image Search Results


    Transport pH profiling of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS. Twenty-four hours later, DOX was added at 10 ng/ml. After 48 h, transport was measured over 2 min at 37 °C with [ 3 H]MTX (0.5 µM) in MBS (20 mM MES, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 5.5, 6.0 and 6.5) and HBS (20 mM Hepes, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 6.8, 7.0, 7.2 and 7.4). The dishes were washed (3×) with ice-cold PBS. The cells were solubilized in 0.5 N NaOH, and the radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity is calculated in units of pmol [ 3 H]MTX per mg of cell protein and results are presented as mean values plus/minus SDs (Panel A, B) or relative values (relative to the maximum activity for each transporter) as mean plus/minus SDs (Panel C, D) from at least 3 experiments. ( A ) Net transport activities of R1-11/Tet-on-RFC/PCFT cells (in the presence of DOX) are shown over a range of pHs. ( B ) The results depict transport activities of R1-11/Tet-on-RFC/PCFT cells at pH 7.2 (optimum for RFC transport) in the absence or presence of 10 μM PT523 (left panel), and at pH 5.5 (optimum for PCFT transport) in the absence or presence of 10 μM AGF94 (right panel). ( C ) RFC transport activities of R1-11/Tet-on-RFC cells (with DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of PCFT specific inhibitor AGF94 of 10 μM; right panel) are shown over a range of pHs. ( D ) PCFT transport activities of R1-11/Tet-on-RFC/PCFT cells (without DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of RFC specific inhibitor PT523 at 10 μM; right panel) are shown over a range of pHs.

    Journal: Scientific Reports

    Article Title: Folate transporter dynamics and therapy with classic and tumor-targeted antifolates

    doi: 10.1038/s41598-021-85818-x

    Figure Lengend Snippet: Transport pH profiling of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS. Twenty-four hours later, DOX was added at 10 ng/ml. After 48 h, transport was measured over 2 min at 37 °C with [ 3 H]MTX (0.5 µM) in MBS (20 mM MES, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 5.5, 6.0 and 6.5) and HBS (20 mM Hepes, 140 mM NaCl, 5 mM KCl, 2 mM MgCl 2 , and 5 mM glucose, for pH 6.8, 7.0, 7.2 and 7.4). The dishes were washed (3×) with ice-cold PBS. The cells were solubilized in 0.5 N NaOH, and the radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity is calculated in units of pmol [ 3 H]MTX per mg of cell protein and results are presented as mean values plus/minus SDs (Panel A, B) or relative values (relative to the maximum activity for each transporter) as mean plus/minus SDs (Panel C, D) from at least 3 experiments. ( A ) Net transport activities of R1-11/Tet-on-RFC/PCFT cells (in the presence of DOX) are shown over a range of pHs. ( B ) The results depict transport activities of R1-11/Tet-on-RFC/PCFT cells at pH 7.2 (optimum for RFC transport) in the absence or presence of 10 μM PT523 (left panel), and at pH 5.5 (optimum for PCFT transport) in the absence or presence of 10 μM AGF94 (right panel). ( C ) RFC transport activities of R1-11/Tet-on-RFC cells (with DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of PCFT specific inhibitor AGF94 of 10 μM; right panel) are shown over a range of pHs. ( D ) PCFT transport activities of R1-11/Tet-on-RFC/PCFT cells (without DOX; left panel) and of R1-11/Tet-on-RFC/PCFT cells (with DOX and in the presence of RFC specific inhibitor PT523 at 10 μM; right panel) are shown over a range of pHs.

    Article Snippet: After 48 h, cells were washed with PBS and disrupted in 10 mM Tris–HCl (pH 7) in the presence of cOmplete Protease Inhibitor Cocktail (Roche Diagnostics, Indianapolis, IN) by sonication.

    Techniques: Radioactivity, Activity Assay

    Characterization of R1-11/Tet-on-FRα single and R1-11/Tet-on-FRα/PCFT double transfectant models. R1-11/Tet-on-FRα or R1-11/Tet-on-FRα/PCFT cells were plated in 60-mm dishes in complete folate-free (FF) RPMI 1640 medium containing 10% fetal bovine serum (FBS) for transport/binding and protein expression assays. Twenty-four hours later, a range of DOX (0, 1, 2.5, 5, 7.5, 10, 25, 50, and 1000 ng/ml) was added. After 48 h, FRα and/or PCFT protein levels for the R1-11/Tet-on-FRα single ( A ) or R1-11/Tet-on-FRα/PCFT double ( B ) models were measured in crude plasma membranes by SDS-PAGE and Western blotting with a HA monoclonal antibody (upper panels) followed by stripping and re-probing with Na + /K + ATPase monoclonal antibody (lower panels) as a loading control. Blots were cropped as needed. The full blots are included in the Supplement (Figs. S2 – S5 ). The molecular mass markers for SDS-PAGE are noted. Densitometry was performed using Odyssey software, and FRα or PCFT protein levels were normalized to those for Na + /K + ATPase and expressed relative to the level at the maximum concentration of DOX. Densitometry results are summarized below the individual lanes and are presented as mean values plus/minus standard deviations (SDs; in parenthesis) from at least 3 experiments. FRα levels ( C , D ) were also determined with [ 3 H]FA at 0 °C for 15 min; PCFT uptake ( F ) was measured with [ 3 H]MTX at pH 5.5 at 37 °C for 2 min. Results are presented as mean values plus/minus SDs from at least 3 experiments. The statistical significance of PCFT transport activities between samples with and without DOX was analyzed by an unpaired t test. An asterisk indicates a statistically significant difference between the mean transport values ( p

    Journal: Scientific Reports

    Article Title: Folate transporter dynamics and therapy with classic and tumor-targeted antifolates

    doi: 10.1038/s41598-021-85818-x

    Figure Lengend Snippet: Characterization of R1-11/Tet-on-FRα single and R1-11/Tet-on-FRα/PCFT double transfectant models. R1-11/Tet-on-FRα or R1-11/Tet-on-FRα/PCFT cells were plated in 60-mm dishes in complete folate-free (FF) RPMI 1640 medium containing 10% fetal bovine serum (FBS) for transport/binding and protein expression assays. Twenty-four hours later, a range of DOX (0, 1, 2.5, 5, 7.5, 10, 25, 50, and 1000 ng/ml) was added. After 48 h, FRα and/or PCFT protein levels for the R1-11/Tet-on-FRα single ( A ) or R1-11/Tet-on-FRα/PCFT double ( B ) models were measured in crude plasma membranes by SDS-PAGE and Western blotting with a HA monoclonal antibody (upper panels) followed by stripping and re-probing with Na + /K + ATPase monoclonal antibody (lower panels) as a loading control. Blots were cropped as needed. The full blots are included in the Supplement (Figs. S2 – S5 ). The molecular mass markers for SDS-PAGE are noted. Densitometry was performed using Odyssey software, and FRα or PCFT protein levels were normalized to those for Na + /K + ATPase and expressed relative to the level at the maximum concentration of DOX. Densitometry results are summarized below the individual lanes and are presented as mean values plus/minus standard deviations (SDs; in parenthesis) from at least 3 experiments. FRα levels ( C , D ) were also determined with [ 3 H]FA at 0 °C for 15 min; PCFT uptake ( F ) was measured with [ 3 H]MTX at pH 5.5 at 37 °C for 2 min. Results are presented as mean values plus/minus SDs from at least 3 experiments. The statistical significance of PCFT transport activities between samples with and without DOX was analyzed by an unpaired t test. An asterisk indicates a statistically significant difference between the mean transport values ( p

    Article Snippet: After 48 h, cells were washed with PBS and disrupted in 10 mM Tris–HCl (pH 7) in the presence of cOmplete Protease Inhibitor Cocktail (Roche Diagnostics, Indianapolis, IN) by sonication.

    Techniques: Transfection, Binding Assay, Expressing, SDS Page, Western Blot, Stripping Membranes, Software, Concentration Assay

    Characterization of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double transfectant models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS for transport and protein expression assays. Twenty-four hours later, a range of DOX (0, 1, 2.5, 5, 7.5, 10, 25, 50, and 1000 ng/ml) was added. After 48 h, RFC and PCFT protein levels for the R1-11/Tet-on-RFC ( A ) and R1-11/Tet-on-RFC/PCFT models (after deglycosylation; “dgRFC” and “dgPCFT” designate the deglycosylated forms of these proteins) ( B ) were measured in crude plasma membranes by SDS-PAGE and Western blotting with HA monoclonal antibody (upper panels), followed by stripping and re-probing with Na + /K + ATPase monoclonal antibody (lower panels) as a loading control. Blots were cropped as needed. The full blots are included in the Supplement (Figs. S6 to S9 ). The molecular mass markers for SDS-PAGE are noted. Densitometry was performed using the Odyssey software, and RFC or PCFT protein levels were normalized to Na + /K + ATPase and expressed relative to the level at the maximum concentration of DOX. Densitometry results are noted below the individual lanes and are presented as mean values plus/minus SDs from at least 3 experiments. RFC ( C , D ) and PCFT ( F ) transport activities were measured with [ 3 H]MTX at 37 °C for 2 min, at pH 7.2 and pH 5.5, respectively. Results are presented as mean values plus/minus SDs from at least 3 experiments. Statistical significance of PCFT transport activities between samples with and without DOX was analyzed by the unpaired t test. An asterisk indicates a statistically significant difference between the mean values of PCFT transport ( p

    Journal: Scientific Reports

    Article Title: Folate transporter dynamics and therapy with classic and tumor-targeted antifolates

    doi: 10.1038/s41598-021-85818-x

    Figure Lengend Snippet: Characterization of R1-11/Tet-on-RFC single and R1-11/Tet-on-RFC/PCFT double transfectant models. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS for transport and protein expression assays. Twenty-four hours later, a range of DOX (0, 1, 2.5, 5, 7.5, 10, 25, 50, and 1000 ng/ml) was added. After 48 h, RFC and PCFT protein levels for the R1-11/Tet-on-RFC ( A ) and R1-11/Tet-on-RFC/PCFT models (after deglycosylation; “dgRFC” and “dgPCFT” designate the deglycosylated forms of these proteins) ( B ) were measured in crude plasma membranes by SDS-PAGE and Western blotting with HA monoclonal antibody (upper panels), followed by stripping and re-probing with Na + /K + ATPase monoclonal antibody (lower panels) as a loading control. Blots were cropped as needed. The full blots are included in the Supplement (Figs. S6 to S9 ). The molecular mass markers for SDS-PAGE are noted. Densitometry was performed using the Odyssey software, and RFC or PCFT protein levels were normalized to Na + /K + ATPase and expressed relative to the level at the maximum concentration of DOX. Densitometry results are noted below the individual lanes and are presented as mean values plus/minus SDs from at least 3 experiments. RFC ( C , D ) and PCFT ( F ) transport activities were measured with [ 3 H]MTX at 37 °C for 2 min, at pH 7.2 and pH 5.5, respectively. Results are presented as mean values plus/minus SDs from at least 3 experiments. Statistical significance of PCFT transport activities between samples with and without DOX was analyzed by the unpaired t test. An asterisk indicates a statistically significant difference between the mean values of PCFT transport ( p

    Article Snippet: After 48 h, cells were washed with PBS and disrupted in 10 mM Tris–HCl (pH 7) in the presence of cOmplete Protease Inhibitor Cocktail (Roche Diagnostics, Indianapolis, IN) by sonication.

    Techniques: Transfection, Expressing, SDS Page, Western Blot, Stripping Membranes, Software, Concentration Assay

    3 H -LCV accumulation by R1-11/Tet-on-FRα, R1-11/Tet-on-FRα/PCFT, R1-11/Tet-on-RFC and R1-11/Tet-on-RFC/PCFT cell line models. R1-11/Tet-on-FRα, R1-11/Tet-on-FRα/PCFT, R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS for 24–48 h. The media was then replaced with complete FF RPMI 1640 medium (pH 6.5, 6.8 or 7.2) containing 10% dialyzed FBS, 25 nM [ 3 H]LCV and a range of DOX (0, 1, 2.5, 5, 10, and 1000 ng/ml). Forty-eight hours later, the dishes were washed (2×) with acid buffer (10 mM sodium acetate, 150 mM NaCl, pH 3.5; for R1-11/Tet-on-FRα or R1-11/Tet-on-FRα/PCFT cells) and 3× with ice-cold PBS. The R1-11/Tet-on-RFC and R1-11/Tet-on-RFC/PCFT cells were washed 3× with ice-cold PBS only. The washed cells were solubilized in 0.5 N NaOH and radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity was calculated in units of pmol [ 3 H]LCV per mg of cell protein. The [ 3 H]LCV uptake results are presented as mean values plus/minus SDs from at least 3 experiments for incubations at pH 6.5 ( A , D ), pH 6.8 ( B , E ) and pH 7.2 ( C , F ). Statistical significance of [ 3 H]LCV uptake values between the single and double transfectants at each DOX dosage was analyzed by an unpaired t test. An asterisk indicates a statistically significant difference between the mean values of [ 3 H]LCV accumulation by single and double transfected cells at each DOX concentration (* p

    Journal: Scientific Reports

    Article Title: Folate transporter dynamics and therapy with classic and tumor-targeted antifolates

    doi: 10.1038/s41598-021-85818-x

    Figure Lengend Snippet: 3 H -LCV accumulation by R1-11/Tet-on-FRα, R1-11/Tet-on-FRα/PCFT, R1-11/Tet-on-RFC and R1-11/Tet-on-RFC/PCFT cell line models. R1-11/Tet-on-FRα, R1-11/Tet-on-FRα/PCFT, R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 60-mm dishes in complete FF RPMI 1640 medium containing 10% FBS for 24–48 h. The media was then replaced with complete FF RPMI 1640 medium (pH 6.5, 6.8 or 7.2) containing 10% dialyzed FBS, 25 nM [ 3 H]LCV and a range of DOX (0, 1, 2.5, 5, 10, and 1000 ng/ml). Forty-eight hours later, the dishes were washed (2×) with acid buffer (10 mM sodium acetate, 150 mM NaCl, pH 3.5; for R1-11/Tet-on-FRα or R1-11/Tet-on-FRα/PCFT cells) and 3× with ice-cold PBS. The R1-11/Tet-on-RFC and R1-11/Tet-on-RFC/PCFT cells were washed 3× with ice-cold PBS only. The washed cells were solubilized in 0.5 N NaOH and radioactive contents and protein concentrations of the alkaline cell homogenates were determined. Intracellular radioactivity was calculated in units of pmol [ 3 H]LCV per mg of cell protein. The [ 3 H]LCV uptake results are presented as mean values plus/minus SDs from at least 3 experiments for incubations at pH 6.5 ( A , D ), pH 6.8 ( B , E ) and pH 7.2 ( C , F ). Statistical significance of [ 3 H]LCV uptake values between the single and double transfectants at each DOX dosage was analyzed by an unpaired t test. An asterisk indicates a statistically significant difference between the mean values of [ 3 H]LCV accumulation by single and double transfected cells at each DOX concentration (* p

    Article Snippet: After 48 h, cells were washed with PBS and disrupted in 10 mM Tris–HCl (pH 7) in the presence of cOmplete Protease Inhibitor Cocktail (Roche Diagnostics, Indianapolis, IN) by sonication.

    Techniques: Radioactivity, Transfection, Concentration Assay

    Impact of RFC/PCFT redundancy on drug sensitivity. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 96-well culture plates (4000 cells/well; 200 μl/well) with complete FF RPMI 1640 including 10% dialyzed FBS supplemented with 25 nM LCV. Drugs with different transport specificities were added, with concentrations from 1 to 1000 nM for AGF102 ( A ), MTX ( B ), PT523 ( C ), PMX ( D ), and AGF94 ( E ). Cells were treated over a range of DOX concentrations (0, 1, 2.5, 5, 10 and 1000 ng/ml) and incubated from 96 to120 h (depending on the growth of the cell models) at 37 °C in a CO 2 incubator. Cell viabilities were measured with a fluorescence-based viability assay (CellTiter-Blue; Promega) and a fluorescence plate reader (emission at 590 nm, excitation at 560 nm) for calculating the drug concentrations that inhibit growth by 50% (IC 50 ). Results are shown as mean IC 50 values + /− SDs from 4 to 15 separate experiments.

    Journal: Scientific Reports

    Article Title: Folate transporter dynamics and therapy with classic and tumor-targeted antifolates

    doi: 10.1038/s41598-021-85818-x

    Figure Lengend Snippet: Impact of RFC/PCFT redundancy on drug sensitivity. R1-11/Tet-on-RFC or R1-11/Tet-on-RFC/PCFT cells were plated in 96-well culture plates (4000 cells/well; 200 μl/well) with complete FF RPMI 1640 including 10% dialyzed FBS supplemented with 25 nM LCV. Drugs with different transport specificities were added, with concentrations from 1 to 1000 nM for AGF102 ( A ), MTX ( B ), PT523 ( C ), PMX ( D ), and AGF94 ( E ). Cells were treated over a range of DOX concentrations (0, 1, 2.5, 5, 10 and 1000 ng/ml) and incubated from 96 to120 h (depending on the growth of the cell models) at 37 °C in a CO 2 incubator. Cell viabilities were measured with a fluorescence-based viability assay (CellTiter-Blue; Promega) and a fluorescence plate reader (emission at 590 nm, excitation at 560 nm) for calculating the drug concentrations that inhibit growth by 50% (IC 50 ). Results are shown as mean IC 50 values + /− SDs from 4 to 15 separate experiments.

    Article Snippet: After 48 h, cells were washed with PBS and disrupted in 10 mM Tris–HCl (pH 7) in the presence of cOmplete Protease Inhibitor Cocktail (Roche Diagnostics, Indianapolis, IN) by sonication.

    Techniques: Incubation, Fluorescence, Viability Assay