Structured Review

Bio-Rad alamarblue
Nonspecific cytotoxicity in MSCs at a variety of siRNA concentrations and charge ratios delivered via polymer diblock or DharmaFECT. (A) MSCs were treated with siRNA at different concentrations with charge ratio of 4:1 (diblock copolymer carrier) or by manufacturer’s recommendations (DharmaFECT) and after 24 hours analyzed for cell survivability using the <t>alamarBlue</t> metabolic assay. (B) MSCs were treated with siRNA at 37.5 nM with charge ratios of 1:1–8:1. Data are from three independent experiments conducted in triplicate relative to survivability of untreated cells (No Treatment) with error bars representing standard deviation. Statistical significance was evaluated at a level of p
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1) Product Images from "Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA"

Article Title: Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA

Journal: Biomacromolecules

doi: 10.1021/bm301294n

Nonspecific cytotoxicity in MSCs at a variety of siRNA concentrations and charge ratios delivered via polymer diblock or DharmaFECT. (A) MSCs were treated with siRNA at different concentrations with charge ratio of 4:1 (diblock copolymer carrier) or by manufacturer’s recommendations (DharmaFECT) and after 24 hours analyzed for cell survivability using the alamarBlue metabolic assay. (B) MSCs were treated with siRNA at 37.5 nM with charge ratios of 1:1–8:1. Data are from three independent experiments conducted in triplicate relative to survivability of untreated cells (No Treatment) with error bars representing standard deviation. Statistical significance was evaluated at a level of p
Figure Legend Snippet: Nonspecific cytotoxicity in MSCs at a variety of siRNA concentrations and charge ratios delivered via polymer diblock or DharmaFECT. (A) MSCs were treated with siRNA at different concentrations with charge ratio of 4:1 (diblock copolymer carrier) or by manufacturer’s recommendations (DharmaFECT) and after 24 hours analyzed for cell survivability using the alamarBlue metabolic assay. (B) MSCs were treated with siRNA at 37.5 nM with charge ratios of 1:1–8:1. Data are from three independent experiments conducted in triplicate relative to survivability of untreated cells (No Treatment) with error bars representing standard deviation. Statistical significance was evaluated at a level of p

Techniques Used: Metabolic Assay, Standard Deviation

2) Product Images from "Platelets and Plasma Stimulate Sheep Rotator Cuff Tendon Tenocytes When Cultured in an Extracellular Matrix Scaffold"

Article Title: Platelets and Plasma Stimulate Sheep Rotator Cuff Tendon Tenocytes When Cultured in an Extracellular Matrix Scaffold

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

doi: 10.1002/jor.23058

Proportion of AlamarBlue reduced (n=6 for all groups) over the course of the experiment. Error bars represent standard error. * = significant difference (P
Figure Legend Snippet: Proportion of AlamarBlue reduced (n=6 for all groups) over the course of the experiment. Error bars represent standard error. * = significant difference (P

Techniques Used:

3) Product Images from "The Putative Guanine Nucleotide Exchange Factor RicA Mediates Upstream Signaling for Growth and Development in Aspergillus"

Article Title: The Putative Guanine Nucleotide Exchange Factor RicA Mediates Upstream Signaling for Growth and Development in Aspergillus

Journal: Eukaryotic Cell

doi: 10.1128/EC.00255-12

Phenotypes caused by ΔAf ricA . (A) WT (AF293), ΔAf ricA (FNJ12), and complemented (C′ Af ricA ; FNJ13) strains were point inoculated on solid MMG plus 0.1% YE and incubated at 37°C for 5 days. Photographs of colony sizes and close-up views of the centers of the colonies and conidiophores are shown. Black arrows indicate abnormal conidiophores at the center of the ΔAf ricA colony. (B) Germination of WT (AF293) and ΔAf ricA (FNJ12) conidia inoculated on MMG plus 0.1% YE plates and incubated for 6 and 12 h. Black arrows in the image for ΔAf ricA indicate germinated conidia at 12 h. (C) Accumulation of brlA , abaA , wetA , and γ-actin mRNA post-asexual developmental induction of WT (AF293) and ΔAf ricA (FNJ12) strains. Development at 0 h indicates vegetative growth in MMG plus 0.1% YE liquid for 18 h. The A. fumigatus ) was used as a control. (D) alamarBlue reduction data, indicating relative cell death rates. The mycelial aggregates of WT, ΔAf ricA , and complemented strains were mixed with the AB reagent to check the cell viability for 7 days. (E) Dry weights of WT, ΔAf ricA , and the complemented strain in MMG plus 0.1% YE submerged cultures were quantified for 7 days at 37°C, 220 rpm.
Figure Legend Snippet: Phenotypes caused by ΔAf ricA . (A) WT (AF293), ΔAf ricA (FNJ12), and complemented (C′ Af ricA ; FNJ13) strains were point inoculated on solid MMG plus 0.1% YE and incubated at 37°C for 5 days. Photographs of colony sizes and close-up views of the centers of the colonies and conidiophores are shown. Black arrows indicate abnormal conidiophores at the center of the ΔAf ricA colony. (B) Germination of WT (AF293) and ΔAf ricA (FNJ12) conidia inoculated on MMG plus 0.1% YE plates and incubated for 6 and 12 h. Black arrows in the image for ΔAf ricA indicate germinated conidia at 12 h. (C) Accumulation of brlA , abaA , wetA , and γ-actin mRNA post-asexual developmental induction of WT (AF293) and ΔAf ricA (FNJ12) strains. Development at 0 h indicates vegetative growth in MMG plus 0.1% YE liquid for 18 h. The A. fumigatus ) was used as a control. (D) alamarBlue reduction data, indicating relative cell death rates. The mycelial aggregates of WT, ΔAf ricA , and complemented strains were mixed with the AB reagent to check the cell viability for 7 days. (E) Dry weights of WT, ΔAf ricA , and the complemented strain in MMG plus 0.1% YE submerged cultures were quantified for 7 days at 37°C, 220 rpm.

Techniques Used: Incubation

4) Product Images from "Chemically Modified DNA Aptamers Bind Interleukin-6 with High Affinity and Inhibit Signaling by Blocking Its Interaction with Interleukin-6 Receptor"

Article Title: Chemically Modified DNA Aptamers Bind Interleukin-6 with High Affinity and Inhibit Signaling by Blocking Its Interaction with Interleukin-6 Receptor

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.532580

Inhibition of tumor cell proliferation by SL1026, SL1033, and tocilizumab. A, inhibition of U266B1 (human myeloma) cell proliferation. Cell proliferation was measured with AlamarBlue, and percent IL-6 inhibition (relative to a no-inhibitor control sample) was plotted. Each bar represents the mean ± S.E. of three experiments. The data were analyzed by one-way analysis of variance (Dunnett's two-tailed), and each data point was compared with control data without IL-6 inhibitor. Statistically significant differences are denoted as *, p value
Figure Legend Snippet: Inhibition of tumor cell proliferation by SL1026, SL1033, and tocilizumab. A, inhibition of U266B1 (human myeloma) cell proliferation. Cell proliferation was measured with AlamarBlue, and percent IL-6 inhibition (relative to a no-inhibitor control sample) was plotted. Each bar represents the mean ± S.E. of three experiments. The data were analyzed by one-way analysis of variance (Dunnett's two-tailed), and each data point was compared with control data without IL-6 inhibitor. Statistically significant differences are denoted as *, p value

Techniques Used: Inhibition, Two Tailed Test

5) Product Images from "The Influence of Pathological Mutations and Proline Substitutions in TDP-43 Glycine-Rich Peptides on Its Amyloid Properties and Cellular Toxicity"

Article Title: The Influence of Pathological Mutations and Proline Substitutions in TDP-43 Glycine-Rich Peptides on Its Amyloid Properties and Cellular Toxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0103644

TDP-43 pathological and glycine to proline mutant peptides exhibit different neurotoxicity. (A) The comparison of cell viability in the presence of D1, G294V, G295S, GGG294PPP, and GGG308PPP for 72 hours determined by AlamarBlue assay. Results were means ± SEM of three independent experiments (* = p
Figure Legend Snippet: TDP-43 pathological and glycine to proline mutant peptides exhibit different neurotoxicity. (A) The comparison of cell viability in the presence of D1, G294V, G295S, GGG294PPP, and GGG308PPP for 72 hours determined by AlamarBlue assay. Results were means ± SEM of three independent experiments (* = p

Techniques Used: Mutagenesis, Alamar Blue Assay

6) Product Images from "Novel Screen to Assess Bactericidal Activity of Compounds Against Non-replicating Mycobacterium abscessus"

Article Title: Novel Screen to Assess Bactericidal Activity of Compounds Against Non-replicating Mycobacterium abscessus

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.02417

Niclosamide has potent activity against non-replicating bacteria. Nutrient-starved bacteria were exposed to nicolsamide for 72 h. alamarBlue ® was added, and RFU measured after 24 h. RFUs were normalized to the DMSO control to express % viability. Data are mean ± SD of four independent biological replicates.
Figure Legend Snippet: Niclosamide has potent activity against non-replicating bacteria. Nutrient-starved bacteria were exposed to nicolsamide for 72 h. alamarBlue ® was added, and RFU measured after 24 h. RFUs were normalized to the DMSO control to express % viability. Data are mean ± SD of four independent biological replicates.

Techniques Used: Activity Assay

Metabolic activity correlates with bactericidal activity. Nutrient-starved bacteria were exposed to compounds for 24–72 h, alamarBlue ® was added and RFU measured after 24 h. Aliquots from the same samples were used to determine: (A) Bacterial viability (CFU; data are the average ± SD of 2 biological replicates); (B) metabolic activity.
Figure Legend Snippet: Metabolic activity correlates with bactericidal activity. Nutrient-starved bacteria were exposed to compounds for 24–72 h, alamarBlue ® was added and RFU measured after 24 h. Aliquots from the same samples were used to determine: (A) Bacterial viability (CFU; data are the average ± SD of 2 biological replicates); (B) metabolic activity.

Techniques Used: Activity Assay

7) Product Images from "A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling"

Article Title: A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling

Journal: PLoS ONE

doi: 10.1371/journal.pone.0001823

The viability of BaFiso cell lines in the absence of IL-3. (A) Parental Ba/F3 derived BY and BC cells, and the BaFiso cell lines BYA and BCS were maintained in the presence or absence of IL-3 (+IL3 and −IL3). Photos were taken 24 h after transferring the cells to medium without IL-3 or in the presence of 3 ng/ml of the recombinant cytokine. (B) Measurement of cell viability using the Alamar blue assay. Alamar blue fluoresces and changes color in response to chemical reduction, and the extent of the conversion is a reflection of cell viability. Metabolic conversion of Alamar blue to its reduced, pink derivative upon cytokine-deprivation (−IL-3) or in its presence (+IL-3). (C), Bar graph showing the results of Alamar Blue cell viability assay. Maximal absorbance of the reduced and oxidized forms of AlamarBlue™, 570 and 600 nm was measured using Victor 1420 multilabel counter 24 h after IL-3 withdrawal. The percentage of cell survival was calculated compared with control cells in the presence of 3 ng/ml of IL3. The data represents three independent experiments performed in triplicate samples. (D) Time course of cell viability upon IL-3 withdrawal. Cells were washed twice in PBS and seeded in media lacking IL-3. Viability was assessed at 12 hour intervals by trypan blue exclusions followed by cell countings. Black rhombs and open squares represent percentage viability of BY and BC cells, respectively. Open triangles and black circles represent percentage viability of BYA and BCS cells, respectively. Data are presented as mean±SD from three independent experiments.
Figure Legend Snippet: The viability of BaFiso cell lines in the absence of IL-3. (A) Parental Ba/F3 derived BY and BC cells, and the BaFiso cell lines BYA and BCS were maintained in the presence or absence of IL-3 (+IL3 and −IL3). Photos were taken 24 h after transferring the cells to medium without IL-3 or in the presence of 3 ng/ml of the recombinant cytokine. (B) Measurement of cell viability using the Alamar blue assay. Alamar blue fluoresces and changes color in response to chemical reduction, and the extent of the conversion is a reflection of cell viability. Metabolic conversion of Alamar blue to its reduced, pink derivative upon cytokine-deprivation (−IL-3) or in its presence (+IL-3). (C), Bar graph showing the results of Alamar Blue cell viability assay. Maximal absorbance of the reduced and oxidized forms of AlamarBlue™, 570 and 600 nm was measured using Victor 1420 multilabel counter 24 h after IL-3 withdrawal. The percentage of cell survival was calculated compared with control cells in the presence of 3 ng/ml of IL3. The data represents three independent experiments performed in triplicate samples. (D) Time course of cell viability upon IL-3 withdrawal. Cells were washed twice in PBS and seeded in media lacking IL-3. Viability was assessed at 12 hour intervals by trypan blue exclusions followed by cell countings. Black rhombs and open squares represent percentage viability of BY and BC cells, respectively. Open triangles and black circles represent percentage viability of BYA and BCS cells, respectively. Data are presented as mean±SD from three independent experiments.

Techniques Used: Derivative Assay, Transferring, Recombinant, Alamar Blue Assay, Viability Assay

8) Product Images from "Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival"

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

Journal: Hepatology Communications

doi: 10.1002/hep4.1181

Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).
Figure Legend Snippet: Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

Techniques Used: Western Blot, Infection, shRNA, Alamar Blue Assay, Fluorescence

Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).
Figure Legend Snippet: Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

Techniques Used: Western Blot, Alamar Blue Assay, Infection, shRNA, Cell Culture, Staining, Fluorescence

9) Product Images from "Rifampicin and rifabutin resistance in 1003 Mycobacterium tuberculosis clinical isolates"

Article Title: Rifampicin and rifabutin resistance in 1003 Mycobacterium tuberculosis clinical isolates

Journal: Journal of Antimicrobial Chemotherapy

doi: 10.1093/jac/dkz048

The rpoB D435V mutation increases the MIC of both rifampicin and rifabutin. Measurement of antibiotic resistance by alamarBlue reduction assay with varying concentrations of rifampicin (a) and rifabutin (b). OD 570 is normalized with the growth of an antibiotic-free control set to 1. Each data point represents the mean of three independent cultures with the standard deviation.
Figure Legend Snippet: The rpoB D435V mutation increases the MIC of both rifampicin and rifabutin. Measurement of antibiotic resistance by alamarBlue reduction assay with varying concentrations of rifampicin (a) and rifabutin (b). OD 570 is normalized with the growth of an antibiotic-free control set to 1. Each data point represents the mean of three independent cultures with the standard deviation.

Techniques Used: Mutagenesis, Standard Deviation

10) Product Images from "Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival"

Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

Journal: Hepatology Communications

doi: 10.1002/hep4.1181

Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).
Figure Legend Snippet: Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

Techniques Used: Western Blot, Infection, shRNA, Alamar Blue Assay, Fluorescence

Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).
Figure Legend Snippet: Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

Techniques Used: Western Blot, Alamar Blue Assay, Infection, shRNA, Cell Culture, Staining, Fluorescence

11) Product Images from "Therapeutic efficacy of liposomal Grb2 antisense oligodeoxynucleotide (L-Grb2) in preclinical models of ovarian and uterine cancer"

Article Title: Therapeutic efficacy of liposomal Grb2 antisense oligodeoxynucleotide (L-Grb2) in preclinical models of ovarian and uterine cancer

Journal: Oncotarget

doi: 10.18632/oncotarget.27667

Effect of Grb2 downregulation on ovarian cancer cell lines. ( A ) Western blot analysis of Grb2 expression in a panel of ovarian cancer cell lines compared with that in the non-transformed epithelial ovarian cell line HIO180. The adjoining graph shows their expression compared with that in HIO180. ( B ) Western blot analysis of Grb2 expression in OVCAR8 cells 72 hours after siGrb2 (Grb2 siRNA) cells compared with that in untreated (UT) and siControl (Con siRNA) cells. ( C ) OVCAR8, SKOV3ip1, A2780, HEYA8, and OVCAR5 cell lines were transfected with siGrb2 or siControl at increasing concentrations. An alamarBlue assay of the cells was performed 72 hours after transfection to determine their percent viability, which is shown in the graphs. The data represent averages of triplicate measurements. ( D ) The percent viability of the five cell lines shown in panel C, 72 hours after transfection of siGrb2 and siControl at 160 nM. ( E ) Western blot analysis of erbB2 expression in a panel of ovarian cancer cell lines compared to fallopian tubal epithelium (FTE). ( F ) Results of an annexin V assay performed to determine the number of apoptotic untreated (UT), siControl-transfected (100 nM), and siGrb2-transfected (100 nM) ovarian cancer cells. The assay was performed 72 hours after transfection. Error bars, SEM. All statistical tests were two-sided. Asterisk indicates statistical significance of ** p
Figure Legend Snippet: Effect of Grb2 downregulation on ovarian cancer cell lines. ( A ) Western blot analysis of Grb2 expression in a panel of ovarian cancer cell lines compared with that in the non-transformed epithelial ovarian cell line HIO180. The adjoining graph shows their expression compared with that in HIO180. ( B ) Western blot analysis of Grb2 expression in OVCAR8 cells 72 hours after siGrb2 (Grb2 siRNA) cells compared with that in untreated (UT) and siControl (Con siRNA) cells. ( C ) OVCAR8, SKOV3ip1, A2780, HEYA8, and OVCAR5 cell lines were transfected with siGrb2 or siControl at increasing concentrations. An alamarBlue assay of the cells was performed 72 hours after transfection to determine their percent viability, which is shown in the graphs. The data represent averages of triplicate measurements. ( D ) The percent viability of the five cell lines shown in panel C, 72 hours after transfection of siGrb2 and siControl at 160 nM. ( E ) Western blot analysis of erbB2 expression in a panel of ovarian cancer cell lines compared to fallopian tubal epithelium (FTE). ( F ) Results of an annexin V assay performed to determine the number of apoptotic untreated (UT), siControl-transfected (100 nM), and siGrb2-transfected (100 nM) ovarian cancer cells. The assay was performed 72 hours after transfection. Error bars, SEM. All statistical tests were two-sided. Asterisk indicates statistical significance of ** p

Techniques Used: Western Blot, Expressing, Transformation Assay, Transfection, Alamar Blue Assay, Annexin V Assay

12) Product Images from "Preparation and Biological Properties of Ring-Substituted Naphthalene-1-Carboxanilides"

Article Title: Preparation and Biological Properties of Ring-Substituted Naphthalene-1-Carboxanilides

Journal: Molecules

doi: 10.3390/molecules190710386

Viability of M. avium subsp. paratuberculosis expressed as percent reduction of alamarBlue over a period of five days after treatment with selected studied compounds 2b , 3c , and 4b in comparison with untreated control of M. avium subsp. paratuberculosis and standards ciprofloxacin (CPX) and rifampicin (RIF) at their MICs ( A ). Viability of M. avium subsp. paratuberculosis expressed as percent reduction of alamarBlue over a period of five days after treatment with selected studied compounds 2b ( B ), 3c ( C ), and 4b ( D ) at different concentrations (1,000–15 µg/mL) in comparison with CPX and RIF at their MICs.
Figure Legend Snippet: Viability of M. avium subsp. paratuberculosis expressed as percent reduction of alamarBlue over a period of five days after treatment with selected studied compounds 2b , 3c , and 4b in comparison with untreated control of M. avium subsp. paratuberculosis and standards ciprofloxacin (CPX) and rifampicin (RIF) at their MICs ( A ). Viability of M. avium subsp. paratuberculosis expressed as percent reduction of alamarBlue over a period of five days after treatment with selected studied compounds 2b ( B ), 3c ( C ), and 4b ( D ) at different concentrations (1,000–15 µg/mL) in comparison with CPX and RIF at their MICs.

Techniques Used:

13) Product Images from "Ebselen Alters Mitochondrial Physiology and Reduces Viability of Rat Hippocampal Astrocytes"

Article Title: Ebselen Alters Mitochondrial Physiology and Reduces Viability of Rat Hippocampal Astrocytes

Journal: DNA and Cell Biology

doi: 10.1089/dna.2012.1939

Analysis of viability of rat hippocampal astrocytes. Cell proliferation and cytotoxicity under the different treatments applied were analyzed studying AlamarBlue ® reduction, as described in Materials and Methods. (A) Incubation of cells in the
Figure Legend Snippet: Analysis of viability of rat hippocampal astrocytes. Cell proliferation and cytotoxicity under the different treatments applied were analyzed studying AlamarBlue ® reduction, as described in Materials and Methods. (A) Incubation of cells in the

Techniques Used: Incubation

14) Product Images from "Cellular Protein Kinase D Modulators Play a Role during Multiple Steps of Herpes Simplex Virus 1 Egress"

Article Title: Cellular Protein Kinase D Modulators Play a Role during Multiple Steps of Herpes Simplex Virus 1 Egress

Journal: Journal of Virology

doi: 10.1128/JVI.01486-18

Impacts of PKD modulators on HSV-1 yields. (A, B) 143B cells were transfected for 48 h with the LipoJet reagent alone or with different siRNAs and dsiRNAs before being infected for 24 h. The supernatants (A) and cell fractions (B) were then harvested, and the virus produced quantified in plaque assays. (C) The viability of 143B cells treated in parallel as described above was measured using alamarBlue. The mean values and SEM from five independent experiments each performed in duplicate are shown in all panels. The data are normalized to the average obtained with samples transfected with LipoJet alone. Significant differences between the results for cells subjected to RNAi and those treated with LipoJet alone were evaluated with Student’s bilateral tests (*, P
Figure Legend Snippet: Impacts of PKD modulators on HSV-1 yields. (A, B) 143B cells were transfected for 48 h with the LipoJet reagent alone or with different siRNAs and dsiRNAs before being infected for 24 h. The supernatants (A) and cell fractions (B) were then harvested, and the virus produced quantified in plaque assays. (C) The viability of 143B cells treated in parallel as described above was measured using alamarBlue. The mean values and SEM from five independent experiments each performed in duplicate are shown in all panels. The data are normalized to the average obtained with samples transfected with LipoJet alone. Significant differences between the results for cells subjected to RNAi and those treated with LipoJet alone were evaluated with Student’s bilateral tests (*, P

Techniques Used: Transfection, Infection, Produced

CERT overexpression hinders viral production. (A) 143B cells were transfected for 24 h with the LipoD293 reagent alone or plasmid expressing hemagglutinin (HA)-tagged wild-type CERT (pCERT). Cells transfected for 24 h were subsequently infected for 24 h, and the supernatants harvested for plaque assays. The mean values and SEM are derived from two independent experiments each performed in duplicate. (B) Cells treated in parallel in 96-well plates were monitored for their viability using alamarBlue. In this case, the bars and error bars show the mean values and SEM from three independent experiments each performed in triplicate. In all cases, the data are normalized to the average value obtained with samples transfected with LipoD293 alone. Student’s bilateral tests showed the significant differences between results for cells transfected with pCERT and cells treated with LipoD293 alone (**, P
Figure Legend Snippet: CERT overexpression hinders viral production. (A) 143B cells were transfected for 24 h with the LipoD293 reagent alone or plasmid expressing hemagglutinin (HA)-tagged wild-type CERT (pCERT). Cells transfected for 24 h were subsequently infected for 24 h, and the supernatants harvested for plaque assays. The mean values and SEM are derived from two independent experiments each performed in duplicate. (B) Cells treated in parallel in 96-well plates were monitored for their viability using alamarBlue. In this case, the bars and error bars show the mean values and SEM from three independent experiments each performed in triplicate. In all cases, the data are normalized to the average value obtained with samples transfected with LipoD293 alone. Student’s bilateral tests showed the significant differences between results for cells transfected with pCERT and cells treated with LipoD293 alone (**, P

Techniques Used: Over Expression, Transfection, Plasmid Preparation, Expressing, Infection, Derivative Assay

Inhibition of CERT with the HPA-12 drug does not affect the egress of the virus. (A) 143B cells were infected in the presence of DMSO or 2.5 μM HPA-12 for 16 h. The supernatants were then harvested and used for plaque assays. The mean values and SEM from three independent experiments performed in duplicate are shown. (B) 143B cells were treated in parallel with 2.5 μM HPA-12 for 16 h, and their viability measured using alamarBlue. Error bars show the SEM of three independent experiments performed in triplicate. In both panels, the data are normalized to the average value obtained with samples treated with DMSO alone. Student’s bilateral tests did not hint at any significant differences between the results obtained using HPA-12 and DMSO alone.
Figure Legend Snippet: Inhibition of CERT with the HPA-12 drug does not affect the egress of the virus. (A) 143B cells were infected in the presence of DMSO or 2.5 μM HPA-12 for 16 h. The supernatants were then harvested and used for plaque assays. The mean values and SEM from three independent experiments performed in duplicate are shown. (B) 143B cells were treated in parallel with 2.5 μM HPA-12 for 16 h, and their viability measured using alamarBlue. Error bars show the SEM of three independent experiments performed in triplicate. In both panels, the data are normalized to the average value obtained with samples treated with DMSO alone. Student’s bilateral tests did not hint at any significant differences between the results obtained using HPA-12 and DMSO alone.

Techniques Used: Inhibition, Infection

15) Product Images from "Anticancer activity of extracts derived from the mature roots of Scutellaria baicalensis on human malignant brain tumor cells"

Article Title: Anticancer activity of extracts derived from the mature roots of Scutellaria baicalensis on human malignant brain tumor cells

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/1472-6882-6-27

The cytotoxic effects of Scutellaria baicalensis extract on normal glial cells and glioma cells. Wells were essentially confluent at the beginning of the experiment. (A) Metabolic activity of normal glial cells (HJ) and cells from a recurrent tumor (MER) were measured using the AlamarBlue™ metabolic assay. Glioma cells were cultured with 100 μg/ml of the S. baicalensis extract for approximately 48 hours. Results were presented as a percentage of the control (untreated). Data show means ± SD of 3 replicates. (B) Photomicrographs of cells 48 hours following treatment with 100 μg/ml of S. baicalensis extract.
Figure Legend Snippet: The cytotoxic effects of Scutellaria baicalensis extract on normal glial cells and glioma cells. Wells were essentially confluent at the beginning of the experiment. (A) Metabolic activity of normal glial cells (HJ) and cells from a recurrent tumor (MER) were measured using the AlamarBlue™ metabolic assay. Glioma cells were cultured with 100 μg/ml of the S. baicalensis extract for approximately 48 hours. Results were presented as a percentage of the control (untreated). Data show means ± SD of 3 replicates. (B) Photomicrographs of cells 48 hours following treatment with 100 μg/ml of S. baicalensis extract.

Techniques Used: Activity Assay, Metabolic Assay, Cell Culture

Time course treatment of ME series cells with S. baicalensis extract. Metabolic activity was measured using AlamarBlue™. Wells were subconfluent on Day 0. Cells were left untreated (■-■), mock treated with 0.4% ethanol (∇-∇) or treated with a 0.4% ethanol solution containing 100 μg/ml S. baicalensis extract (△-△). Data is mean ± SD using 5 replicates. (A) cells from primary tumor (ME), (B) ME cells selected for resistance to 10 μg/ml BCNU (ME drug), (C) cells from recurrent tumor (MER), (D) cells from recurrent tumor selected for resistance to 10 μg/ml BCNU (MER drug).
Figure Legend Snippet: Time course treatment of ME series cells with S. baicalensis extract. Metabolic activity was measured using AlamarBlue™. Wells were subconfluent on Day 0. Cells were left untreated (■-■), mock treated with 0.4% ethanol (∇-∇) or treated with a 0.4% ethanol solution containing 100 μg/ml S. baicalensis extract (△-△). Data is mean ± SD using 5 replicates. (A) cells from primary tumor (ME), (B) ME cells selected for resistance to 10 μg/ml BCNU (ME drug), (C) cells from recurrent tumor (MER), (D) cells from recurrent tumor selected for resistance to 10 μg/ml BCNU (MER drug).

Techniques Used: Activity Assay

16) Product Images from "Mesenchymal Stem Cell Secretome Enhancement by Nicotinamide and Vasoactive Intestinal Peptide: A New Therapeutic Approach for Retinal Degenerative Diseases"

Article Title: Mesenchymal Stem Cell Secretome Enhancement by Nicotinamide and Vasoactive Intestinal Peptide: A New Therapeutic Approach for Retinal Degenerative Diseases

Journal: Stem Cells International

doi: 10.1155/2020/9463548

Cell proliferation in glucose oxidase stressed ARPE-19 cells cultured with the pharmaceutical compositions (PhC). Damaged ARPE-19 cell proliferation was measured using alamarBlue® at days 1, 3, and 6 of cell culture with the PhC. (a) aMSC-CM combined (PhC 1) with NIC, VIP, or NIC+VIP tended to reduce cell proliferation of glucose oxidase stressed ARPE-19 cells. While the effect was greatest in the presence of NIC+VIP, none of the differences were significant ( p > 0.05). (b) aMSC-CM stimulated-combined (PhC 2) and (c) aMSC-CM stimulated (PhC 3) conditions tended to have higher better proliferation rates in glucose oxidase stressed ARPE-19 cells, though again, the differences were not significant. Bars represent the mean of normalized fluorescence unit (NFU) ± SEM. Statistical analysis: repeated measures ANOVA with Tukey's post hoc tests and Bonferroni corrections for multiple testing. Three experiments ( n = 3) in triplicate were performed. aMSC: adipose-derived mesenchymal stem cells; CM: conditioned medium; NIC: nicotinamide; VIP: vasoactive intestinal peptide.
Figure Legend Snippet: Cell proliferation in glucose oxidase stressed ARPE-19 cells cultured with the pharmaceutical compositions (PhC). Damaged ARPE-19 cell proliferation was measured using alamarBlue® at days 1, 3, and 6 of cell culture with the PhC. (a) aMSC-CM combined (PhC 1) with NIC, VIP, or NIC+VIP tended to reduce cell proliferation of glucose oxidase stressed ARPE-19 cells. While the effect was greatest in the presence of NIC+VIP, none of the differences were significant ( p > 0.05). (b) aMSC-CM stimulated-combined (PhC 2) and (c) aMSC-CM stimulated (PhC 3) conditions tended to have higher better proliferation rates in glucose oxidase stressed ARPE-19 cells, though again, the differences were not significant. Bars represent the mean of normalized fluorescence unit (NFU) ± SEM. Statistical analysis: repeated measures ANOVA with Tukey's post hoc tests and Bonferroni corrections for multiple testing. Three experiments ( n = 3) in triplicate were performed. aMSC: adipose-derived mesenchymal stem cells; CM: conditioned medium; NIC: nicotinamide; VIP: vasoactive intestinal peptide.

Techniques Used: Cell Culture, Fluorescence, Derivative Assay

17) Product Images from "Identifying a Neuromedin U Receptor 2 Splice Variant and Determining Its Roles in the Regulation of Signaling and Tumorigenesis In Vitro"

Article Title: Identifying a Neuromedin U Receptor 2 Splice Variant and Determining Its Roles in the Regulation of Signaling and Tumorigenesis In Vitro

Journal: PLoS ONE

doi: 10.1371/journal.pone.0136836

Expression of NMUR2S and its effects on cancer cell signaling and progression. (A) The transcripts of NMU , NMUR1 , NMUR2 and NMUR2S were compared in diverse cancer cell lines. β-actin ( ACTB) levels served as loading controls. (B) SKOV-3 cells overexpressing eGFP or NMUR2S , or (D) THP-1 cells with eGFP or NMUR2S knockdown were then treated with 100 nM NMU for different intervals before estimating the amounts of phosphorylated ERK1/2 (pERK1/2) and total ERK 1/2 by Western blotting. The ratios of pERK1/2 to ERK 1/2 in each sample were then calculated by densitometry and are shown below. (C) Cell proliferation rates between eGFP -expressing and NMUR2S -expressing SKOV-3 cells were compared using the AlamarBlue assay. The fluorescence value of the cells on day 0 (D0) served as a one-fold control. (E) Cell proliferation rates between GFP -knockdown and NMUR2S -knockdown THP-1 cells were compared by counting the cell numbers directly. The amount of THP-1 cells on day 0 (D0) served as a one-fold control. *, P
Figure Legend Snippet: Expression of NMUR2S and its effects on cancer cell signaling and progression. (A) The transcripts of NMU , NMUR1 , NMUR2 and NMUR2S were compared in diverse cancer cell lines. β-actin ( ACTB) levels served as loading controls. (B) SKOV-3 cells overexpressing eGFP or NMUR2S , or (D) THP-1 cells with eGFP or NMUR2S knockdown were then treated with 100 nM NMU for different intervals before estimating the amounts of phosphorylated ERK1/2 (pERK1/2) and total ERK 1/2 by Western blotting. The ratios of pERK1/2 to ERK 1/2 in each sample were then calculated by densitometry and are shown below. (C) Cell proliferation rates between eGFP -expressing and NMUR2S -expressing SKOV-3 cells were compared using the AlamarBlue assay. The fluorescence value of the cells on day 0 (D0) served as a one-fold control. (E) Cell proliferation rates between GFP -knockdown and NMUR2S -knockdown THP-1 cells were compared by counting the cell numbers directly. The amount of THP-1 cells on day 0 (D0) served as a one-fold control. *, P

Techniques Used: Expressing, Western Blot, Alamar Blue Assay, Fluorescence

18) Product Images from "Interferon-γ-modified dendritic cells suppress B cell function and ameliorate the development of experimental autoimmune myasthenia gravis"

Article Title: Interferon-γ-modified dendritic cells suppress B cell function and ameliorate the development of experimental autoimmune myasthenia gravis

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2004.02585.x

IFN- γ -modulated DC did not affect T cell proliferation but reduced the number of nAChR antibody-secreting cells. Lymph nodes and spleens were obtained on day 33 p.i. from four rats from each group. MNC were prepared and cultured in the presence of nAChR or the irrelevant antigen, MBP, for 72 h. (a) Proliferation of MNC from spleen and lymph nodes assessed after addition of AlamarBlue to the cultures according to the manufacturer's protocol. Absorbances were measured after 4 h. (b) Number of nAChR antibody-secreting cells per 10 5 MNC from spleen and lymph nodes. Naive DC = rats injected with naive DC; IFN- γ -DC = rats injected with IFN- γ -modulated DC; IFN- γ +1-MT-DC = rats injected with IFN -γ +1-MT-modulated DC. Columns refer to means and bars to s.e. The difference between the naive DC-treated group and IFN- γ -DC-treated group is represented as * P
Figure Legend Snippet: IFN- γ -modulated DC did not affect T cell proliferation but reduced the number of nAChR antibody-secreting cells. Lymph nodes and spleens were obtained on day 33 p.i. from four rats from each group. MNC were prepared and cultured in the presence of nAChR or the irrelevant antigen, MBP, for 72 h. (a) Proliferation of MNC from spleen and lymph nodes assessed after addition of AlamarBlue to the cultures according to the manufacturer's protocol. Absorbances were measured after 4 h. (b) Number of nAChR antibody-secreting cells per 10 5 MNC from spleen and lymph nodes. Naive DC = rats injected with naive DC; IFN- γ -DC = rats injected with IFN- γ -modulated DC; IFN- γ +1-MT-DC = rats injected with IFN -γ +1-MT-modulated DC. Columns refer to means and bars to s.e. The difference between the naive DC-treated group and IFN- γ -DC-treated group is represented as * P

Techniques Used: Cell Culture, Injection

19) Product Images from "Combination of carbonic anhydrase inhibitor, acetazolamide, and sulforaphane, reduces the viability and growth of bronchial carcinoid cell lines"

Article Title: Combination of carbonic anhydrase inhibitor, acetazolamide, and sulforaphane, reduces the viability and growth of bronchial carcinoid cell lines

Journal: BMC Cancer

doi: 10.1186/1471-2407-13-378

AZ and SFN Treatment Interfere with Exogenous 5-HT Stimulated Growth of Lung Carcinoid Cells: figures (a, b) represent AlamarBlue assay; dose response for AZ (black), SFN (gray) and AZ + SFN (white) treatments of H-727 and H-720 cells (0–80 μM; 7 Days, 205% FBS) after adding 5-HT exogenously into the culture medium, respectively; *(p
Figure Legend Snippet: AZ and SFN Treatment Interfere with Exogenous 5-HT Stimulated Growth of Lung Carcinoid Cells: figures (a, b) represent AlamarBlue assay; dose response for AZ (black), SFN (gray) and AZ + SFN (white) treatments of H-727 and H-720 cells (0–80 μM; 7 Days, 205% FBS) after adding 5-HT exogenously into the culture medium, respectively; *(p

Techniques Used: Alamar Blue Assay

AZ and/or SFN Treatment Inhibit Growth of Lung Carcinoid and Fetal Lung Fibroblast Cells (H-727 and H-720); 48 hours, 7 days) and (FLF): AlamarBlue assay; dose response for AZ, SFN and AZ + SFN; (0-80 μM) treatment in H-727 (a,b and c), H-720 (d, e and f) cells 48 hours (gray) and 7 days (white) and AlamarBlue assay; dose response for AZ (white), SFN (gray) and AZ + SFN (black); (0-80 μM) treatment in FLF (g, h and i) cells, 7 days. The significance level compared to control (p value) was specified as follows: *(p
Figure Legend Snippet: AZ and/or SFN Treatment Inhibit Growth of Lung Carcinoid and Fetal Lung Fibroblast Cells (H-727 and H-720); 48 hours, 7 days) and (FLF): AlamarBlue assay; dose response for AZ, SFN and AZ + SFN; (0-80 μM) treatment in H-727 (a,b and c), H-720 (d, e and f) cells 48 hours (gray) and 7 days (white) and AlamarBlue assay; dose response for AZ (white), SFN (gray) and AZ + SFN (black); (0-80 μM) treatment in FLF (g, h and i) cells, 7 days. The significance level compared to control (p value) was specified as follows: *(p

Techniques Used: Alamar Blue Assay, Significance Assay

20) Product Images from "Interactions of newly synthesized platinum nanoparticles with ICR-191 and their potential application"

Article Title: Interactions of newly synthesized platinum nanoparticles with ICR-191 and their potential application

Journal: Scientific Reports

doi: 10.1038/s41598-019-41092-6

Influence of the platinum nanoparticles (PtNPs) on ICR-191 mutagenic activity in tested eukaryotic cell lines: ( A ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a presence of ICR-191 (4.57–450 µM), based on alamarBlue assay. Data are expressed as the mean ± standard deviation and ( B ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a presence of PtNPs (0.2–40 µL/mL), based on alamarBlue assay. Data are expressed as the mean ± standard deviation and ( C ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a mixture of ICR-191 (45.7 µM) and PtNPs (0.2–40 µL/mL), based on alamarBlue assay. Data are expressed as the mean ± standard deviation, and ( D ) confocal microscopy live analysis of the impact of platinum nanoparticles (PtNPs) on ICR-191 fluorescence in the HaCaT and MelJuSo cell lines. Cells were treated with 45.7 µM ICR-191; treated with 45.7 µM ICR-191 and 3 ng/mL PtNPs mixture, or preincubated with 3 ng/mL PtNPs and subsequently treated with 45.7 µM ICR-191 (PtNPs preincubation). Time of incubation in the presence of ICR-191 indicated above particular panels. *Values significantly different from the amount of alamarBlue reduced by untreated control cells (p
Figure Legend Snippet: Influence of the platinum nanoparticles (PtNPs) on ICR-191 mutagenic activity in tested eukaryotic cell lines: ( A ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a presence of ICR-191 (4.57–450 µM), based on alamarBlue assay. Data are expressed as the mean ± standard deviation and ( B ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a presence of PtNPs (0.2–40 µL/mL), based on alamarBlue assay. Data are expressed as the mean ± standard deviation and ( C ) comparison of the cell viability of HaCaT and MelJuSo cell lines incubated for 72 h in a mixture of ICR-191 (45.7 µM) and PtNPs (0.2–40 µL/mL), based on alamarBlue assay. Data are expressed as the mean ± standard deviation, and ( D ) confocal microscopy live analysis of the impact of platinum nanoparticles (PtNPs) on ICR-191 fluorescence in the HaCaT and MelJuSo cell lines. Cells were treated with 45.7 µM ICR-191; treated with 45.7 µM ICR-191 and 3 ng/mL PtNPs mixture, or preincubated with 3 ng/mL PtNPs and subsequently treated with 45.7 µM ICR-191 (PtNPs preincubation). Time of incubation in the presence of ICR-191 indicated above particular panels. *Values significantly different from the amount of alamarBlue reduced by untreated control cells (p

Techniques Used: Activity Assay, Incubation, Alamar Blue Assay, Standard Deviation, Confocal Microscopy, Fluorescence

21) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

Journal: mSphere

doi: 10.1128/mSphere.00199-17

CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

Techniques Used: Expressing, Incubation, Fluorescence, Software

22) Product Images from "Porous Polyethylene Coated with Functionalized Hydroxyapatite Particles as a Bone Reconstruction Material"

Article Title: Porous Polyethylene Coated with Functionalized Hydroxyapatite Particles as a Bone Reconstruction Material

Journal: Materials

doi: 10.3390/ma11040521

( A ) AlamarBlue ® assay of CL1 cells cultured on porous polyethylene and polyethylene + hydroxylapatite scaffolds; ( B ) CL1 cells grown on a polyethylene scaffold; ( C ) CL2 cells grown on polyethylene + hydroxylapatite scaffolds stained with acridine orange, which stains all nucleated cells green. The magnification used was 10×.
Figure Legend Snippet: ( A ) AlamarBlue ® assay of CL1 cells cultured on porous polyethylene and polyethylene + hydroxylapatite scaffolds; ( B ) CL1 cells grown on a polyethylene scaffold; ( C ) CL2 cells grown on polyethylene + hydroxylapatite scaffolds stained with acridine orange, which stains all nucleated cells green. The magnification used was 10×.

Techniques Used: Alamar Blue Assay, Cell Culture, Staining

23) Product Images from "Characterisation of the expression and function of the GM-CSF receptor ?-chain in mice"

Article Title: Characterisation of the expression and function of the GM-CSF receptor ?-chain in mice

Journal: European Journal of Immunology

doi: 10.1002/eji.200636892

Effect of GM-Fc on Ba/F3 cells expressing the deletion and truncation GMRα isoforms. (A) The binding capacity of GM-Fc to Ba/F3 cells expressing the deletion and truncation GMRα from C57BL/6 mice was determined by FACS as indicated previously. (B) Ba/F3:pFB, Ba/F3:GMRα (C57BL/6), the deletion and truncation isoforms (C57BL/6) were cultured in the presence of either GM-CSF, GM-Fc, or IL-3-conditioned medium or in the absence of cytokine. Cell proliferation was determined using Alamarblue as indicated previously.
Figure Legend Snippet: Effect of GM-Fc on Ba/F3 cells expressing the deletion and truncation GMRα isoforms. (A) The binding capacity of GM-Fc to Ba/F3 cells expressing the deletion and truncation GMRα from C57BL/6 mice was determined by FACS as indicated previously. (B) Ba/F3:pFB, Ba/F3:GMRα (C57BL/6), the deletion and truncation isoforms (C57BL/6) were cultured in the presence of either GM-CSF, GM-Fc, or IL-3-conditioned medium or in the absence of cytokine. Cell proliferation was determined using Alamarblue as indicated previously.

Techniques Used: Expressing, Binding Assay, Mouse Assay, FACS, Cell Culture

Effect of GM-Fc on Ba/F3 cells expressing full-length GMRα isoforms. (A) Ba/F3:pFB (filled squares), Ba/F3:GMRα (Balb/c; open circles), and Ba/F3:GMRα (C57BL/6; filled circles) were cultured in the presence of either GM-CSF or GM-Fc and cell proliferation was determined using Alamarblue as indicated previously. (B) Expressing percentage Alamarblue reduction obtained when GM-Fc or GM-CSF was used as a growth factor as a ratio shows that both agents have a similar capacity to support proliferation of Ba/F3:GMRα cells. (C) The ability of the C57BL/6 and Balb/c isoforms of GMRα to bind to the cytokine was assessed using GM-Fc and analysed by non-linear regression. Data shown represent pooled data from two independent experiments, and error bars shown the SEM.
Figure Legend Snippet: Effect of GM-Fc on Ba/F3 cells expressing full-length GMRα isoforms. (A) Ba/F3:pFB (filled squares), Ba/F3:GMRα (Balb/c; open circles), and Ba/F3:GMRα (C57BL/6; filled circles) were cultured in the presence of either GM-CSF or GM-Fc and cell proliferation was determined using Alamarblue as indicated previously. (B) Expressing percentage Alamarblue reduction obtained when GM-Fc or GM-CSF was used as a growth factor as a ratio shows that both agents have a similar capacity to support proliferation of Ba/F3:GMRα cells. (C) The ability of the C57BL/6 and Balb/c isoforms of GMRα to bind to the cytokine was assessed using GM-Fc and analysed by non-linear regression. Data shown represent pooled data from two independent experiments, and error bars shown the SEM.

Techniques Used: Expressing, Cell Culture

Functionality of polymorphic forms of GMRα. (A) Full-length GMRα from Balb/c and C57BL/6 mice were retrovirally transduced into Ba/F3 cells. Ba/F3 cells transduced with empty vector or GMRα-encoding vectors were cultured in the presence of 4 ng/mL of GM-CSF (filled circles) or IL-3 (open circles) for the indicated number of days (left panels) or in the indicated concentrations of cytokine for 8 days (right panels). Cell proliferation was measured by determining the percentage of reduction of Alamarblue. (B) The ratio of proliferation in response to GM-CSF and IL-3 was calculated to compare the polymorphic variants; filled squares: Ba/F3:pFB; open circles: Ba/F3:GMRα (Balb/c); filled circles: Ba/F3:GMRα (C57BL/6).
Figure Legend Snippet: Functionality of polymorphic forms of GMRα. (A) Full-length GMRα from Balb/c and C57BL/6 mice were retrovirally transduced into Ba/F3 cells. Ba/F3 cells transduced with empty vector or GMRα-encoding vectors were cultured in the presence of 4 ng/mL of GM-CSF (filled circles) or IL-3 (open circles) for the indicated number of days (left panels) or in the indicated concentrations of cytokine for 8 days (right panels). Cell proliferation was measured by determining the percentage of reduction of Alamarblue. (B) The ratio of proliferation in response to GM-CSF and IL-3 was calculated to compare the polymorphic variants; filled squares: Ba/F3:pFB; open circles: Ba/F3:GMRα (Balb/c); filled circles: Ba/F3:GMRα (C57BL/6).

Techniques Used: Mouse Assay, Transduction, Plasmid Preparation, Cell Culture

24) Product Images from "The Effects of Cyclic Hydrostatic Pressure on Chondrogenesis and Viability of Human Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Agarose Constructs"

Article Title: The Effects of Cyclic Hydrostatic Pressure on Chondrogenesis and Viability of Human Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Agarose Constructs

Journal: Tissue Engineering. Part A

doi: 10.1089/ten.tea.2012.0015

Average percent reduction of alamarBlue for hASCs and hMSCs. Different letters represent statistical differences ( p
Figure Legend Snippet: Average percent reduction of alamarBlue for hASCs and hMSCs. Different letters represent statistical differences ( p

Techniques Used:

25) Product Images from "Incorporation of vitamin E in poly(3hydroxybutyrate)/Bioglass composite films: effect on surface properties and cell attachment"

Article Title: Incorporation of vitamin E in poly(3hydroxybutyrate)/Bioglass composite films: effect on surface properties and cell attachment

Journal: Journal of the Royal Society Interface

doi: 10.1098/rsif.2008.0278

Cell proliferation study for 1, 4 and 7 days, using AlamarBlue assay on ( a ) samples with and without vitamin E, ( b ) all tested samples relative to the control (control set to 100%). The data ( n =3; error bars=±s.d.) were compared using Student's t -test and differences were considered significant when * p
Figure Legend Snippet: Cell proliferation study for 1, 4 and 7 days, using AlamarBlue assay on ( a ) samples with and without vitamin E, ( b ) all tested samples relative to the control (control set to 100%). The data ( n =3; error bars=±s.d.) were compared using Student's t -test and differences were considered significant when * p

Techniques Used: Alamar Blue Assay

26) Product Images from "Enhanced trophic factor secretion by mesenchymal stem/stromal cells with Glycine-Histidine-Lysine (GHK)-modified alginate hydrogels"

Article Title: Enhanced trophic factor secretion by mesenchymal stem/stromal cells with Glycine-Histidine-Lysine (GHK)-modified alginate hydrogels

Journal: Acta biomaterialia

doi: 10.1016/j.actbio.2014.01.020

Cell viability of MSC treated with increasing concentration of GHK for 3 days in monolayer culture: (A) metabolic activity assayed by percent reduction of alamarBlue; (B) protein concentration; (C) caspase 3/7 activity for n=4.
Figure Legend Snippet: Cell viability of MSC treated with increasing concentration of GHK for 3 days in monolayer culture: (A) metabolic activity assayed by percent reduction of alamarBlue; (B) protein concentration; (C) caspase 3/7 activity for n=4.

Techniques Used: Concentration Assay, Activity Assay, Protein Concentration

27) Product Images from "Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA"

Article Title: Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA

Journal: Biomacromolecules

doi: 10.1021/bm301294n

Nonspecific cytotoxicity in MSCs at a variety of siRNA concentrations and charge ratios delivered via polymer diblock or DharmaFECT. (A) MSCs were treated with siRNA at different concentrations with charge ratio of 4:1 (diblock copolymer carrier) or by manufacturer’s recommendations (DharmaFECT) and after 24 hours analyzed for cell survivability using the alamarBlue metabolic assay. (B) MSCs were treated with siRNA at 37.5 nM with charge ratios of 1:1–8:1. Data are from three independent experiments conducted in triplicate relative to survivability of untreated cells (No Treatment) with error bars representing standard deviation. Statistical significance was evaluated at a level of p
Figure Legend Snippet: Nonspecific cytotoxicity in MSCs at a variety of siRNA concentrations and charge ratios delivered via polymer diblock or DharmaFECT. (A) MSCs were treated with siRNA at different concentrations with charge ratio of 4:1 (diblock copolymer carrier) or by manufacturer’s recommendations (DharmaFECT) and after 24 hours analyzed for cell survivability using the alamarBlue metabolic assay. (B) MSCs were treated with siRNA at 37.5 nM with charge ratios of 1:1–8:1. Data are from three independent experiments conducted in triplicate relative to survivability of untreated cells (No Treatment) with error bars representing standard deviation. Statistical significance was evaluated at a level of p

Techniques Used: Metabolic Assay, Standard Deviation

28) Product Images from "Design of Experiments Approach to Engineer Cell-Secreted Matrices for Directing Osteogenic Differentiation"

Article Title: Design of Experiments Approach to Engineer Cell-Secreted Matrices for Directing Osteogenic Differentiation

Journal: Annals of Biomedical Engineering

doi: 10.1007/s10439-010-0217-x

hMSC proliferation and viability are enhanced when cultured on DM1 compared to culture on DM2 or control substrates. (a) Total DNA on each substrate at 1, 4, and 7 days post-seeding. (b) AlamarBlue reduction by hMSCs cultured on each substrate at 1, 4, and 7 days. (c) Calcein uptake by hMSCs seeded on each substrate and under environmental stress for 24 h. # p
Figure Legend Snippet: hMSC proliferation and viability are enhanced when cultured on DM1 compared to culture on DM2 or control substrates. (a) Total DNA on each substrate at 1, 4, and 7 days post-seeding. (b) AlamarBlue reduction by hMSCs cultured on each substrate at 1, 4, and 7 days. (c) Calcein uptake by hMSCs seeded on each substrate and under environmental stress for 24 h. # p

Techniques Used: Cell Culture

29) Product Images from "A comparative evaluation of the effect of polymer chemistry and fiber orientation on mesenchymal stem cell differentiation"

Article Title: A comparative evaluation of the effect of polymer chemistry and fiber orientation on mesenchymal stem cell differentiation

Journal: Journal of Biomedical Materials Research. Part a

doi: 10.1002/jbm.a.35829

Cell viability of chondrogenic but not adipogenic and osteogenic cells was influenced by scaffold material. (A) Initial cell attachment of MSCs on scaffolds was assessed by AlamarBlue at day 1 after seeding, RFU = relative fluorescent units. Cell viability of adipogenic (B), chondrogenic (C), and osteogenic (D) MSCs was measured at days 7 and 14 after seeding ( n = 3 MSC donors)
Figure Legend Snippet: Cell viability of chondrogenic but not adipogenic and osteogenic cells was influenced by scaffold material. (A) Initial cell attachment of MSCs on scaffolds was assessed by AlamarBlue at day 1 after seeding, RFU = relative fluorescent units. Cell viability of adipogenic (B), chondrogenic (C), and osteogenic (D) MSCs was measured at days 7 and 14 after seeding ( n = 3 MSC donors)

Techniques Used: Cell Attachment Assay

30) Product Images from "Intrinsic Osteoinductivity of Porous Titanium Scaffold for Bone Tissue Engineering"

Article Title: Intrinsic Osteoinductivity of Porous Titanium Scaffold for Bone Tissue Engineering

Journal: International Journal of Biomaterials

doi: 10.1155/2017/5093063

Biocompatibility and osteoinductivity of porous Ti scaffolds. AlamarBlue absorbance of BMMSCs seeded on HAp coated noncoated scaffolds over 28 days; Alizarin Red absorbance normalized to noncoated samples at day 28. ∗ shows significantly higher value compared to other samples ( p
Figure Legend Snippet: Biocompatibility and osteoinductivity of porous Ti scaffolds. AlamarBlue absorbance of BMMSCs seeded on HAp coated noncoated scaffolds over 28 days; Alizarin Red absorbance normalized to noncoated samples at day 28. ∗ shows significantly higher value compared to other samples ( p

Techniques Used:

31) Product Images from "Functions of innate and acquired immune system are reduced in domestic pigeons (Columba livia domestica) given a low protein diet"

Article Title: Functions of innate and acquired immune system are reduced in domestic pigeons (Columba livia domestica) given a low protein diet

Journal: Royal Society Open Science

doi: 10.1098/rsos.150408

Experiment 2. Lymphocyte proliferation (% increase of mitogen-stimulated cells relative to un-stimulated cells) on y -axis, stimulated by ( a ) ConA at 5 µg ml −1 , ( b ) LPS at 2.5 µg ml −1 and ( c ) PMA at 5 µg ml −1 on pigeons fed the 6, 10 and 14% crude protein (CP) diets on x -axis at 4 (black) and 8 (grey) hour incubation after adding alamarBlue® (mean ± s.e., n = 17: 6 and 10%; n = 16: 14%). Different lower-case letters within the same hour indicate significant differences at p
Figure Legend Snippet: Experiment 2. Lymphocyte proliferation (% increase of mitogen-stimulated cells relative to un-stimulated cells) on y -axis, stimulated by ( a ) ConA at 5 µg ml −1 , ( b ) LPS at 2.5 µg ml −1 and ( c ) PMA at 5 µg ml −1 on pigeons fed the 6, 10 and 14% crude protein (CP) diets on x -axis at 4 (black) and 8 (grey) hour incubation after adding alamarBlue® (mean ± s.e., n = 17: 6 and 10%; n = 16: 14%). Different lower-case letters within the same hour indicate significant differences at p

Techniques Used: Incubation

32) Product Images from "The Effects of Cyclic Hydrostatic Pressure on Chondrogenesis and Viability of Human Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Agarose Constructs"

Article Title: The Effects of Cyclic Hydrostatic Pressure on Chondrogenesis and Viability of Human Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Agarose Constructs

Journal: Tissue Engineering. Part A

doi: 10.1089/ten.tea.2012.0015

Average percent reduction of alamarBlue for hASCs and hMSCs. Different letters represent statistical differences ( p
Figure Legend Snippet: Average percent reduction of alamarBlue for hASCs and hMSCs. Different letters represent statistical differences ( p

Techniques Used:

33) Product Images from "Genome-wide mRNA and miRNA expression profiling reveal multiple regulatory networks in colorectal cancer"

Article Title: Genome-wide mRNA and miRNA expression profiling reveal multiple regulatory networks in colorectal cancer

Journal: Cell Death & Disease

doi: 10.1038/cddis.2014.556

Inhibition of EZH2 using DZNep mediates significant reduction in cell viability and in vitro migration in colon cancer cells. ( a ) HT115, HT-29, and SW620 cells were treated with the indicated dose of DZNep, and cell viability was measured on days 4 and 8 posttreatment using the alamarBlue assay. Data are presented as mean±S.E., n =8. ( b ) DZNep treatment (5 days) led to significant reduction in EZH2 protein expression in HT-29 and SW620 cells. Similarly, DZNep treatment led to substantial reduction in H3K273me in the colon cancer cells. * P
Figure Legend Snippet: Inhibition of EZH2 using DZNep mediates significant reduction in cell viability and in vitro migration in colon cancer cells. ( a ) HT115, HT-29, and SW620 cells were treated with the indicated dose of DZNep, and cell viability was measured on days 4 and 8 posttreatment using the alamarBlue assay. Data are presented as mean±S.E., n =8. ( b ) DZNep treatment (5 days) led to significant reduction in EZH2 protein expression in HT-29 and SW620 cells. Similarly, DZNep treatment led to substantial reduction in H3K273me in the colon cancer cells. * P

Techniques Used: Inhibition, In Vitro, Migration, Alamar Blue Assay, Expressing

34) Product Images from "Naturally Derived Anti-HIV Polysaccharide Peptide (PSP) Triggers a Toll-Like Receptor 4-Dependent Antiviral Immune Response"

Article Title: Naturally Derived Anti-HIV Polysaccharide Peptide (PSP) Triggers a Toll-Like Receptor 4-Dependent Antiviral Immune Response

Journal: Journal of Immunology Research

doi: 10.1155/2018/8741698

Polysaccharide peptide (PSP) activates human cells without significant cytotoxicity. HIV-1-infected and uninfected THP1 cells and freshly isolated human peripheral blood mononuclear cells (PBMC) were exposed twice (treated at day 0 and day 3) with various PSP concentrations (0–1000 μ g/ml) over a six-day period, with 0 μ g/ml as negative control. (a) Half maximal effective concentration (EC50) was determined by measuring secreted embryonic alkaline phosphatase (SEAP) activity in the culture supernatant using QUANTI-Blue assay after 6 days of treatment. Reduction of alamarBlue was used to determine the optimal time course between 3 and 6 days of treatments in either (b) HIV-1-negative and (c) HIV-1-positive THP1 cells. Cell viability was confirmed by tetrazolium MTT assay after a six-day period in (d) HIV−/HIV+ THP1 cells and (e) HIV−/HIV+ human PBMCs. Statistical significance was determined using one-way ANOVA, N = 3 ( ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗∗ p ≤ 0.0001).
Figure Legend Snippet: Polysaccharide peptide (PSP) activates human cells without significant cytotoxicity. HIV-1-infected and uninfected THP1 cells and freshly isolated human peripheral blood mononuclear cells (PBMC) were exposed twice (treated at day 0 and day 3) with various PSP concentrations (0–1000 μ g/ml) over a six-day period, with 0 μ g/ml as negative control. (a) Half maximal effective concentration (EC50) was determined by measuring secreted embryonic alkaline phosphatase (SEAP) activity in the culture supernatant using QUANTI-Blue assay after 6 days of treatment. Reduction of alamarBlue was used to determine the optimal time course between 3 and 6 days of treatments in either (b) HIV-1-negative and (c) HIV-1-positive THP1 cells. Cell viability was confirmed by tetrazolium MTT assay after a six-day period in (d) HIV−/HIV+ THP1 cells and (e) HIV−/HIV+ human PBMCs. Statistical significance was determined using one-way ANOVA, N = 3 ( ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗∗ p ≤ 0.0001).

Techniques Used: Infection, Isolation, Negative Control, Concentration Assay, Activity Assay, MTT Assay

35) Product Images from "The RUNX1/IL-34/CSF-1R axis is an autocrinally regulated modulator of resistance to BRAF-V600E inhibition in melanoma"

Article Title: The RUNX1/IL-34/CSF-1R axis is an autocrinally regulated modulator of resistance to BRAF-V600E inhibition in melanoma

Journal: JCI Insight

doi: 10.1172/jci.insight.120422

Coinhibition of BRAF V600E and CSF-1R is synergistic in melanoma. ( A ) Using a constant ratio, the V600E inhibitor was combined with either the CSF-1R inhibitor or a MEK inhibitor based on the IC 50 value of each drug and used to treat melanoma cells in quadruplicate for 72 hours. The alamarBlue assay was then performed to estimate cell growth inhibition, and curves (mean ± SEM) are shown with the combination index (CI) indicated in the table below for each condition where values
Figure Legend Snippet: Coinhibition of BRAF V600E and CSF-1R is synergistic in melanoma. ( A ) Using a constant ratio, the V600E inhibitor was combined with either the CSF-1R inhibitor or a MEK inhibitor based on the IC 50 value of each drug and used to treat melanoma cells in quadruplicate for 72 hours. The alamarBlue assay was then performed to estimate cell growth inhibition, and curves (mean ± SEM) are shown with the combination index (CI) indicated in the table below for each condition where values

Techniques Used: Alamar Blue Assay, Inhibition

36) Product Images from "Shiga Toxin Mediated Neurologic Changes in Murine Model of Disease"

Article Title: Shiga Toxin Mediated Neurologic Changes in Murine Model of Disease

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2016.00114

Metabolic activity of Stx-treated microvascular endothelial cells . bEnd.3 immortalized mouse cerebral cortex microvascular endothelial cells (A) , primary human cerebral cortex microvascular endothelial cells (B) , primary human neonatal dermal microvascular endothelial cells (C) , and CDC.HMEC-1 immortalized human dermal microvascular endothelial cells (D) were incubated with Stx1 (solid lines) or Stx2a (dashed lines) for 42 h. The toxin containing media was removed and fresh media containing 10% alamarBlue was added. Cells were incubated for an additional 3 h and the fluorescent reduction of alamarBlue was measured every 30 min. The 1 h time point is shown except for subconfluent BMECs which depicts the 3 h time point. Graphs depict toxin-treated cells as a percent of untreated control cells. Results are the average of three individual experiments and error bars correspond to standard deviation of the mean. TNF-α upregulates surface ICAM-1 (E) . Human brain endothelial cells were incubated with 10 ng/ml TNF-α for 24 h, stained for surface expression of ICAM-1 (CD54) and analyzed by FACS.
Figure Legend Snippet: Metabolic activity of Stx-treated microvascular endothelial cells . bEnd.3 immortalized mouse cerebral cortex microvascular endothelial cells (A) , primary human cerebral cortex microvascular endothelial cells (B) , primary human neonatal dermal microvascular endothelial cells (C) , and CDC.HMEC-1 immortalized human dermal microvascular endothelial cells (D) were incubated with Stx1 (solid lines) or Stx2a (dashed lines) for 42 h. The toxin containing media was removed and fresh media containing 10% alamarBlue was added. Cells were incubated for an additional 3 h and the fluorescent reduction of alamarBlue was measured every 30 min. The 1 h time point is shown except for subconfluent BMECs which depicts the 3 h time point. Graphs depict toxin-treated cells as a percent of untreated control cells. Results are the average of three individual experiments and error bars correspond to standard deviation of the mean. TNF-α upregulates surface ICAM-1 (E) . Human brain endothelial cells were incubated with 10 ng/ml TNF-α for 24 h, stained for surface expression of ICAM-1 (CD54) and analyzed by FACS.

Techniques Used: Activity Assay, Incubation, Standard Deviation, Staining, Expressing, FACS

37) Product Images from "Extended Release Combination Antibiotic Therapy from a Bone Void Filling Putty for Treatment of Osteomyelitis"

Article Title: Extended Release Combination Antibiotic Therapy from a Bone Void Filling Putty for Treatment of Osteomyelitis

Journal: Pharmaceutics

doi: 10.3390/pharmaceutics11110592

The alamarBlue cell viability assay showed a significant difference in cell viability between the control and the treatment groups with day 1, week 1 and week 3 drug release milieu ( p
Figure Legend Snippet: The alamarBlue cell viability assay showed a significant difference in cell viability between the control and the treatment groups with day 1, week 1 and week 3 drug release milieu ( p

Techniques Used: Viability Assay

38) Product Images from "Analysis of Virion-Incorporated Host Proteins Required for Herpes Simplex Virus Type 1 Infection through a RNA Interference Screen"

Article Title: Analysis of Virion-Incorporated Host Proteins Required for Herpes Simplex Virus Type 1 Infection through a RNA Interference Screen

Journal: PLoS ONE

doi: 10.1371/journal.pone.0053276

siRNA screen against the host proteins identified in mature extracellular HSV-1 virions. 143B cells were transfected for 48 hours with Lipofectamine alone or with siRNA duplexes targeting the indicated cellular proteins (gene names are indicated on the left). Forty-eight hours later, cellular viability was assessed by alamarBlue or cells were infected in parallel with K26GFP at a MOI of 5 for a further 24 hours. The fluorescence in the supernatant was quantified using a spectrofluorometer. The data was normalized to the mean value obtained with Lipofectamine only samples. Bilateral Student's T-tests were performed to detect significant hits compared to the Lipofectamine only control (*: p
Figure Legend Snippet: siRNA screen against the host proteins identified in mature extracellular HSV-1 virions. 143B cells were transfected for 48 hours with Lipofectamine alone or with siRNA duplexes targeting the indicated cellular proteins (gene names are indicated on the left). Forty-eight hours later, cellular viability was assessed by alamarBlue or cells were infected in parallel with K26GFP at a MOI of 5 for a further 24 hours. The fluorescence in the supernatant was quantified using a spectrofluorometer. The data was normalized to the mean value obtained with Lipofectamine only samples. Bilateral Student's T-tests were performed to detect significant hits compared to the Lipofectamine only control (*: p

Techniques Used: Transfection, Infection, Fluorescence

Screening method. A ) 143B cells were seeded in 24-well plates 24 hours prior to transfection. Cells were then transfected with siRNA and incubated 48 hours before being infected with HSV-1 K26GFP for an additional 24 hours. Supernatants were collected and their fluorescence was measured using a Gemini EM spectrofluorometer. As a cytotoxicity control, cell viability was assessed in parallel using alamarBlue 2 days post-transfection. In addition, the cells were lysed and used for Westerns to validate the siRNA knockdowns. B ) The fluorescence of pre-titered infectious HSV-1 K26GFP particles serially diluted in PBS was quantified using a spectrofluorometer. The fluorescence obtained was linear with the titers through the entire selection (r 2 = 0.9968), thus demonstrating the advanced sensitivity of the device. The gray area denotes the typical of values obtained in the RNA interference screen below. RFU: relative fluorescence units.
Figure Legend Snippet: Screening method. A ) 143B cells were seeded in 24-well plates 24 hours prior to transfection. Cells were then transfected with siRNA and incubated 48 hours before being infected with HSV-1 K26GFP for an additional 24 hours. Supernatants were collected and their fluorescence was measured using a Gemini EM spectrofluorometer. As a cytotoxicity control, cell viability was assessed in parallel using alamarBlue 2 days post-transfection. In addition, the cells were lysed and used for Westerns to validate the siRNA knockdowns. B ) The fluorescence of pre-titered infectious HSV-1 K26GFP particles serially diluted in PBS was quantified using a spectrofluorometer. The fluorescence obtained was linear with the titers through the entire selection (r 2 = 0.9968), thus demonstrating the advanced sensitivity of the device. The gray area denotes the typical of values obtained in the RNA interference screen below. RFU: relative fluorescence units.

Techniques Used: Transfection, Incubation, Infection, Fluorescence, Selection

39) Product Images from "NMU signaling promotes endometrial cancer cell progression by modulating adhesion signaling"

Article Title: NMU signaling promotes endometrial cancer cell progression by modulating adhesion signaling

Journal: Oncotarget

doi: 10.18632/oncotarget.7169

Matrigel culture remedies the effects of NMU knockdown in RL95-2 cells RL95-2 cells without or with NMU knockdown were seeded onto the Matrigel-coated dishes for culture. ( A ) The growth rates were compared using the AlamarBlue assay. The fluorescence value on day 0 (D0) served as the one-fold control. The results are shown as the mean ± SD. n.s, no significance. ( B ) Represented images of cell morphology were shown on Day 5 of culture. Scale bar, 100 μm. ( C ) In the upper panel, the levels of phospho-SRC protein in GFP-knockdown control or NMU -knockdown cells seeded on the standard or Matrigel-coated dishes were compared by Western blotting on Day 5 of culture. Total SRC protein served as the internal control. In the lower panel, the band intensities were further quantified by densitometer in three individual experiments. Fold changes in the phospho-SRC/total SRC level between cells seeded on the standard and Matrigel-coated dishes were normalized and compared. Data are shown as the mean ± SEM. * p
Figure Legend Snippet: Matrigel culture remedies the effects of NMU knockdown in RL95-2 cells RL95-2 cells without or with NMU knockdown were seeded onto the Matrigel-coated dishes for culture. ( A ) The growth rates were compared using the AlamarBlue assay. The fluorescence value on day 0 (D0) served as the one-fold control. The results are shown as the mean ± SD. n.s, no significance. ( B ) Represented images of cell morphology were shown on Day 5 of culture. Scale bar, 100 μm. ( C ) In the upper panel, the levels of phospho-SRC protein in GFP-knockdown control or NMU -knockdown cells seeded on the standard or Matrigel-coated dishes were compared by Western blotting on Day 5 of culture. Total SRC protein served as the internal control. In the lower panel, the band intensities were further quantified by densitometer in three individual experiments. Fold changes in the phospho-SRC/total SRC level between cells seeded on the standard and Matrigel-coated dishes were normalized and compared. Data are shown as the mean ± SEM. * p

Techniques Used: Alamar Blue Assay, Fluorescence, Western Blot

Effects of NMU knockdown on the cell growth and morphology of endometrial cancer cells ( A ) Ishikawa and ( B ) RL95-2 cells with or without NMU knockdown were cultured in the growth medium. ( C ) HEC1A cells with eGFP- or NMUR2 -overexpression were cultured in the growth medium supplemented without or with 100 nM NMU. The growth rates were compared using the AlamarBlue assay. The fluorescence value of the cells on day 0 (D0) served as the one-fold control. Data are shown as the mean ± SD. * p
Figure Legend Snippet: Effects of NMU knockdown on the cell growth and morphology of endometrial cancer cells ( A ) Ishikawa and ( B ) RL95-2 cells with or without NMU knockdown were cultured in the growth medium. ( C ) HEC1A cells with eGFP- or NMUR2 -overexpression were cultured in the growth medium supplemented without or with 100 nM NMU. The growth rates were compared using the AlamarBlue assay. The fluorescence value of the cells on day 0 (D0) served as the one-fold control. Data are shown as the mean ± SD. * p

Techniques Used: Cell Culture, Over Expression, Alamar Blue Assay, Fluorescence

40) Product Images from "ROS and ERK1/2-mediated caspase-9 activation increases XAF1 expression in dexamethasone-induced apoptosis of EBV-transformed B cells"

Article Title: ROS and ERK1/2-mediated caspase-9 activation increases XAF1 expression in dexamethasone-induced apoptosis of EBV-transformed B cells

Journal: International Journal of Oncology

doi: 10.3892/ijo.2013.1949

Dex induced apoptosis in a dose- and time-dependent manner in EBV-transformed B cells. (A) EBV-transformed B cells (5×10 4 cells/well) were cultured in 96-well plates. After 24 h, cell proliferation was measured by AlamarBlue assay. RFU is the relative fluorescence unit. (B and C) EBV-transformed B cells and (D) PBMCs were treated with 10, 50, 100 and 200 μ M of Dex for 2, 4, 8, 16 and 24 h. The percentage of apoptotic cells was estimated by Annexin V/7-AAD staining. Dot plot graphs show percentage of viable cells (Annexin V − /7-AAD − ), early-phage apoptotic cells (Annexin-V + /7-AAD − ), late-phage apoptotic cells (Annexin-V + /7-AAD + ), and necrotic cells (Annexin-V − /7-AAD + ). To measure disruption of Δψ m , cells were stained with DiOC 6 . Diminished DiOC 6 fluorescence indicates Δψ m disruption and percentages indicates the cell proportion in each bar. Results are representative of three independent experiments.
Figure Legend Snippet: Dex induced apoptosis in a dose- and time-dependent manner in EBV-transformed B cells. (A) EBV-transformed B cells (5×10 4 cells/well) were cultured in 96-well plates. After 24 h, cell proliferation was measured by AlamarBlue assay. RFU is the relative fluorescence unit. (B and C) EBV-transformed B cells and (D) PBMCs were treated with 10, 50, 100 and 200 μ M of Dex for 2, 4, 8, 16 and 24 h. The percentage of apoptotic cells was estimated by Annexin V/7-AAD staining. Dot plot graphs show percentage of viable cells (Annexin V − /7-AAD − ), early-phage apoptotic cells (Annexin-V + /7-AAD − ), late-phage apoptotic cells (Annexin-V + /7-AAD + ), and necrotic cells (Annexin-V − /7-AAD + ). To measure disruption of Δψ m , cells were stained with DiOC 6 . Diminished DiOC 6 fluorescence indicates Δψ m disruption and percentages indicates the cell proportion in each bar. Results are representative of three independent experiments.

Techniques Used: Transformation Assay, Cell Culture, Alamar Blue Assay, Fluorescence, Staining

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Metabolic Assay:

Article Title: Controlling mesenchymal stem cell gene expression using polymer-mediated delivery of siRNA
Article Snippet: .. Forty-eight hours after treatment, an alamarBlue (AdB Serotec) metabolic assay for cell viability was performed and the data is summarized in . ..

Concentration Assay:

Article Title: A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling
Article Snippet: .. AlamarBlue™ (Serotec, Oxford, UK) was added to the culture medium at a final concentration of 10% (v-v) and after 24 hours, absorbance was measured at the two different wavelengths of maximal absorbance of the reduced and oxidized forms of AlamarBlue™, 570 and 600 nm using Victor 1420 Multilabel Counter (Perkin-Elmer, Wellesley, USA). ..

Incubation:

Article Title: Chemically Modified DNA Aptamers Bind Interleukin-6 with High Affinity and Inhibit Signaling by Blocking Its Interaction with Interleukin-6 Receptor
Article Snippet: .. AlamarBlue (Bio-Rad, catalog no. BUF012A) was added, and cells were incubated an additional 2–3 h at 37 °C. .. Fluorescence (excitation at 560 nm and emission at 590 nm) was measured with a luminometer (Wallac 1420 ARVO Light, PerkinElmer Life Sciences).

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    Bio-Rad alamarblue colorimetric test
    Viability of peripheral blood mononuclear cells (PBMC) from healthy subjects (NI, n = 12) after 72-h in vitro treatment with reference drug BZ and K777, PYR, and FUR anti- T. cruzi lead compounds by <t>alamarBlue.</t> Data are presented as means ± standard errors of the means (SEM). Significance is indicated by asterisks: P
    Alamarblue Colorimetric Test, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad alamarblue
    Inhibition of tumor cell proliferation by SL1026, SL1033, and tocilizumab. A, inhibition of U266B1 (human myeloma) cell proliferation. Cell proliferation was measured with <t>AlamarBlue,</t> and percent IL-6 inhibition (relative to a no-inhibitor control sample) was plotted. Each bar represents the mean ± S.E. of three experiments. The data were analyzed by one-way analysis of variance (Dunnett's two-tailed), and each data point was compared with control data without IL-6 inhibitor. Statistically significant differences are denoted as *, p value
    Alamarblue, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Viability of peripheral blood mononuclear cells (PBMC) from healthy subjects (NI, n = 12) after 72-h in vitro treatment with reference drug BZ and K777, PYR, and FUR anti- T. cruzi lead compounds by alamarBlue. Data are presented as means ± standard errors of the means (SEM). Significance is indicated by asterisks: P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Identification of Anti- Trypanosoma cruzi Lead Compounds with Putative Immunomodulatory Activity

    doi: 10.1128/AAC.01834-17

    Figure Lengend Snippet: Viability of peripheral blood mononuclear cells (PBMC) from healthy subjects (NI, n = 12) after 72-h in vitro treatment with reference drug BZ and K777, PYR, and FUR anti- T. cruzi lead compounds by alamarBlue. Data are presented as means ± standard errors of the means (SEM). Significance is indicated by asterisks: P

    Article Snippet: The measurement of cytotoxicity of the compounds of interest by the viability and proliferation of PBMC was performed from NI subject cells by the alamarBlue colorimetric test (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: In Vitro

    Inhibition of tumor cell proliferation by SL1026, SL1033, and tocilizumab. A, inhibition of U266B1 (human myeloma) cell proliferation. Cell proliferation was measured with AlamarBlue, and percent IL-6 inhibition (relative to a no-inhibitor control sample) was plotted. Each bar represents the mean ± S.E. of three experiments. The data were analyzed by one-way analysis of variance (Dunnett's two-tailed), and each data point was compared with control data without IL-6 inhibitor. Statistically significant differences are denoted as *, p value

    Journal: The Journal of Biological Chemistry

    Article Title: Chemically Modified DNA Aptamers Bind Interleukin-6 with High Affinity and Inhibit Signaling by Blocking Its Interaction with Interleukin-6 Receptor

    doi: 10.1074/jbc.M113.532580

    Figure Lengend Snippet: Inhibition of tumor cell proliferation by SL1026, SL1033, and tocilizumab. A, inhibition of U266B1 (human myeloma) cell proliferation. Cell proliferation was measured with AlamarBlue, and percent IL-6 inhibition (relative to a no-inhibitor control sample) was plotted. Each bar represents the mean ± S.E. of three experiments. The data were analyzed by one-way analysis of variance (Dunnett's two-tailed), and each data point was compared with control data without IL-6 inhibitor. Statistically significant differences are denoted as *, p value

    Article Snippet: AlamarBlue (Bio-Rad, catalog no. BUF012A) was added, and cells were incubated an additional 2–3 h at 37 °C.

    Techniques: Inhibition, Two Tailed Test

    Niclosamide has potent activity against non-replicating bacteria. Nutrient-starved bacteria were exposed to nicolsamide for 72 h. alamarBlue ® was added, and RFU measured after 24 h. RFUs were normalized to the DMSO control to express % viability. Data are mean ± SD of four independent biological replicates.

    Journal: Frontiers in Microbiology

    Article Title: Novel Screen to Assess Bactericidal Activity of Compounds Against Non-replicating Mycobacterium abscessus

    doi: 10.3389/fmicb.2018.02417

    Figure Lengend Snippet: Niclosamide has potent activity against non-replicating bacteria. Nutrient-starved bacteria were exposed to nicolsamide for 72 h. alamarBlue ® was added, and RFU measured after 24 h. RFUs were normalized to the DMSO control to express % viability. Data are mean ± SD of four independent biological replicates.

    Article Snippet: Assay plates were incubated in a humidified incubator for 48 h at 37°C after which 50 μL of 20% v/v alamarBlue® in PBS-Tyl (Bio-Rad) was added using a Multidrop Combi.

    Techniques: Activity Assay

    Metabolic activity correlates with bactericidal activity. Nutrient-starved bacteria were exposed to compounds for 24–72 h, alamarBlue ® was added and RFU measured after 24 h. Aliquots from the same samples were used to determine: (A) Bacterial viability (CFU; data are the average ± SD of 2 biological replicates); (B) metabolic activity.

    Journal: Frontiers in Microbiology

    Article Title: Novel Screen to Assess Bactericidal Activity of Compounds Against Non-replicating Mycobacterium abscessus

    doi: 10.3389/fmicb.2018.02417

    Figure Lengend Snippet: Metabolic activity correlates with bactericidal activity. Nutrient-starved bacteria were exposed to compounds for 24–72 h, alamarBlue ® was added and RFU measured after 24 h. Aliquots from the same samples were used to determine: (A) Bacterial viability (CFU; data are the average ± SD of 2 biological replicates); (B) metabolic activity.

    Article Snippet: Assay plates were incubated in a humidified incubator for 48 h at 37°C after which 50 μL of 20% v/v alamarBlue® in PBS-Tyl (Bio-Rad) was added using a Multidrop Combi.

    Techniques: Activity Assay

    Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

    Journal: Hepatology Communications

    Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

    doi: 10.1002/hep4.1181

    Figure Lengend Snippet: Knockdown of IGF1R enhanced the inhibitory efficacy of sorafenib in HCC cells by inhibiting AKT. (A) HCC cells were treated with sorafenib (1.25 μM for Hep3B, 2.5 μM for HepG2, and 5 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) Expressions of IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles and then treated with 1.25 μM sorafenib. (D) Expressions of IGF1R, AKT, and GAPDH proteins were examined by western blotting in Hep3B cells infected with scrambled shRNA, IGF1R shRNA, constitutively active AKT, or both lentiviral particles. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles and then treated with 1.25 μM sorafenib. Each experiment was repeated at least 3 times. Abbreviations: GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit; shScr, short hairpin scrambled. Values in C and E were mean ± SD (n = 3 in each group).

    Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

    Techniques: Western Blot, Infection, shRNA, Alamar Blue Assay, Fluorescence

    Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

    Journal: Hepatology Communications

    Article Title: Inhibition of insulin‐like growth factor 1 receptor enhances the efficacy of sorafenib in inhibiting hepatocellular carcinoma cell growth and survival

    doi: 10.1002/hep4.1181

    Figure Lengend Snippet: Ceritinib suppressed HCC cell growth by inhibiting the IGF1R/AKT pathway. (A) HCC cells were treated with ceritinib (0.5 μM for Hep3B, 1 μM for HepG2, and 2 μM for Huh7) for 24 hours. Expressions of p‐IGF1R, IGF1R, p‐AKT (ser473), AKT, p‐ERK, ERK, and GAPDH proteins were examined by western blotting. (B) HCC cells were treated with ceritinib at different doses for 48 hours. Cell proliferation was analyzed by the alamarBlue assay. (C) Cell proliferation was analyzed by the alamarBlue assay in Hep3B cells infected with IGF1R shRNAs and scrambled shRNA lentiviral particles. (D) Hep3B cells infected with control or constitutively active AKT lentiviral particles were treated with 0.5 μM ceritinib for 48 hours. Cells were then cultured for 14 days and stained with 0.5% crystal violet. (E) Cell proliferation was analyzed by the alamarBlue assay in IGF1R knockdown Hep3B cells infected with control or constitutively active AKT lentiviral particles. Each experiment was repeated at least 3 times. Abbreviations: DMSO, dimethyl sulfoxide; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; RFU, relative fluorescence unit. Values in B, C, D, and E were mean ± SD (n = 3 in each group).

    Article Snippet: At different time points, culture media was removed and alamarBlue (BUF012A; Bio‐Rad, Hercules, CA) solution (1:10 dilution in phosphate‐buffered saline) was added to the cells.

    Techniques: Western Blot, Alamar Blue Assay, Infection, shRNA, Cell Culture, Staining, Fluorescence