human cell lines thp 1 (ATCC)


Structured Review

Human Cell Lines Thp 1, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human cell lines thp 1/product/ATCC
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "RUNX3 Regulates Intercellular Adhesion Molecule 3 (ICAM-3) Expression during Macrophage Differentiation and Monocyte Extravasation"
Article Title: RUNX3 Regulates Intercellular Adhesion Molecule 3 (ICAM-3) Expression during Macrophage Differentiation and Monocyte Extravasation
Journal: PLoS ONE
doi: 10.1371/journal.pone.0033313

Figure Legend Snippet: A. Determination of ICAM-3 expression in BLM, Jurkat, K-562 and THP-1 cell lines by Western blot. As a control, β-actin expression levels were also determined. The experiment was performed twice and one of the experiments is shown. B. The ICAM-3 promoter-based constructs pCD50-1000, pCD50-500 and pCD50-200 were transfected in Jurkat (ICAM-3+), THP-1 (ICAM-3+), BLM (ICAM-3−) and K-562 (ICAM-3−) cell lines. After 48 hours cells were lysed and luciferase activity determined. For each reporter construct, promoter activity is expressed relative to the activity produced by the reporter plasmid in BLM cells, arbitrarily set to 1, after normalization for transfection efficiency. Data represent mean ± SD of 4 independent experiments using two different DNA preparations. (*p<0,05 for pCD50-1000Luc in THP-1 and Jurkat and p = 0.8 for K-562 when compared with the activity of pCD50-1000Luc in BLM cells; **p<0.005 for pCD50-500Luc in THP-1 and Jurkat and p = 0.5 for K-562 when compared with the activity of pCD50-500Luc in BLM cells; and * p<0.05 for pCD50-200Luc in THP-1 and Jurkat cells and p = 0.2 for K-562 cells when compared with the activity of pCD50-200Luc in BLM cells). C. Nucleotide sequence of the 5′-regulatory region of the ICAM-3 gene. The transcriptional initiation sites are identified by * and the major transcription initiation site is denoted by +1. First exon nucleotides are shown in boldface type and are underlined. The derived amino acid sequence is shown under the coding region of the first exon. Underlined areas correspond to consensus sequences for RUNX, C/EBP and Ets transcription factors.
Techniques Used: Expressing, Western Blot, Construct, Transfection, Luciferase, Activity Assay, Produced, Plasmid Preparation, Sequencing, Derivative Assay

Figure Legend Snippet: A. EMSA was performed on the indicated oligonucleotides spanning the −157/−14 region of the ICAM-3 promoter using nuclear extracts from THP-1, K-562 and Jurkat cells. The position of the major retarded species is indicated. B. EMSA was performed on the ICAM3.3 and ICAM3.5 oligonucleotides using nuclear extracts from the indicated COS-7 cells transfected with an empty expression vector (pCDNA3) or with either RUNX1 or RUNX3 together with CBF-β expression vector. The position of the RUNX1- and RUNX3-containing complex is shown. C. EMSA was performed on the ICAM3.5 and ICAM3.3 oligonucleotides using nuclear extracts from Jurkat cells in the absence (−) or presence of unlabeled competitor oligonucleotides (ICAM3.5, ICAM3.5mutRUNX, ICAM3.3, ICAM3.3mutRUNX, AMLcons) or polyclonal antisera against CD209 (Control antibody, Cnt Ab) or RUNX1 proteins (R-3034). The position of RUNX1-containing complexes are shown. Unlabeled competitor oligonucleotides were added at a 100-fold molar excess. D. EMSA was performed on the ICAM3.4 oligonucleotide using nuclear extracts from THP-1 cells in the absence (−) or presence of unlabeled competitor oligonucleotides (ICAM3.4, ICAM3.4mutCEBP, C/EBPcons) or polyclonal antibody against CD209 (Control antibody, Cnt Ab) or C/EBPα proteins (α-C/EBPα). The position of C/EBPα-containing complexes are shown. Unlabeled competitor oligonucleotides were added at a 100-fold molar excess. In A–D, EMSA's were performed twice with similar result and a representative experiment is shown. E. ICAM-3 promoter-based oligonucleotides with mutated nucleotides in lowercase and their relative positions. F. In vivo occupancy of the ICAM-3 promoter by RUNX1. Chromatin immunoprecipitation on Jurkat cells was performed with an affinity-purified polyclonal antisera specific for RUNX1 or purified rabbit IgG. Immunoprecipitated chromatin was analyzed by PCR using a pair of ICAM-3 promoter-specific primers that amplify a 234-bp fragment flanking the RUNX-binding sites at −80 and −29. ChIP experiment was performed twice with similar results, and a representative experiment is shown.
Techniques Used: Transfection, Expressing, Plasmid Preparation, In Vivo, Chromatin Immunoprecipitation, Affinity Purification, Purification, Immunoprecipitation, Binding Assay

Figure Legend Snippet: A. Disruption of the RUNX-binding elements leads to increased ICAM-3 gene promoter activity in THP-1 cells. THP-1 cells were transfected with the indicated reporter plasmids and luciferase activity was determined after 24 h. Promoter activity is expressed relative to the activity produced by the wild-type CD50-200Luc, arbitrarily set to 1, after normalization for transfection efficiency. Data represent mean ± SD of 6 independent experiments using two different DNA preparations. B. Knockdown on RUNX3 results in increased ICAM-3 mRNA levels. THP-1 cells were nucleofected with either siRNA for RUNX3 (two different RUNX3 specific-siRNA, siRNA RUNX3#1 and siRNA RUNX3#2) or a control siRNA (siRNA CNT). After 24 hours, total RNA was isolated and ICAM-3 mRNA was measured via quantitative RT-PCR (left). Results are expressed as Relative mRNA levels (relative to GAPDH mRNA levels and the ICAM-3 mRNA level in control siRNA-nucleofected cells). Data represent mean ± SD of 3 independent experiments. To confirm siRNA efficiency, one-fifth of the cells were lysed and underwent western blotting (right). The western blot was performed twice with similar results and one of the experiments is shown.
Techniques Used: Binding Assay, Activity Assay, Transfection, Luciferase, Produced, Isolation, Quantitative RT-PCR, Western Blot
human cell lines thp 1 (ATCC)


Structured Review

Human Cell Lines Thp 1, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human cell lines thp 1/product/ATCC
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "RUNX3 Regulates Intercellular Adhesion Molecule 3 (ICAM-3) Expression during Macrophage Differentiation and Monocyte Extravasation"
Article Title: RUNX3 Regulates Intercellular Adhesion Molecule 3 (ICAM-3) Expression during Macrophage Differentiation and Monocyte Extravasation
Journal: PLoS ONE
doi: 10.1371/journal.pone.0033313

Figure Legend Snippet: A. Determination of ICAM-3 expression in BLM, Jurkat, K-562 and THP-1 cell lines by Western blot. As a control, β-actin expression levels were also determined. The experiment was performed twice and one of the experiments is shown. B. The ICAM-3 promoter-based constructs pCD50-1000, pCD50-500 and pCD50-200 were transfected in Jurkat (ICAM-3+), THP-1 (ICAM-3+), BLM (ICAM-3−) and K-562 (ICAM-3−) cell lines. After 48 hours cells were lysed and luciferase activity determined. For each reporter construct, promoter activity is expressed relative to the activity produced by the reporter plasmid in BLM cells, arbitrarily set to 1, after normalization for transfection efficiency. Data represent mean ± SD of 4 independent experiments using two different DNA preparations. (*p<0,05 for pCD50-1000Luc in THP-1 and Jurkat and p = 0.8 for K-562 when compared with the activity of pCD50-1000Luc in BLM cells; **p<0.005 for pCD50-500Luc in THP-1 and Jurkat and p = 0.5 for K-562 when compared with the activity of pCD50-500Luc in BLM cells; and * p<0.05 for pCD50-200Luc in THP-1 and Jurkat cells and p = 0.2 for K-562 cells when compared with the activity of pCD50-200Luc in BLM cells). C. Nucleotide sequence of the 5′-regulatory region of the ICAM-3 gene. The transcriptional initiation sites are identified by * and the major transcription initiation site is denoted by +1. First exon nucleotides are shown in boldface type and are underlined. The derived amino acid sequence is shown under the coding region of the first exon. Underlined areas correspond to consensus sequences for RUNX, C/EBP and Ets transcription factors.
Techniques Used: Expressing, Western Blot, Construct, Transfection, Luciferase, Activity Assay, Produced, Plasmid Preparation, Sequencing, Derivative Assay

Figure Legend Snippet: A. EMSA was performed on the indicated oligonucleotides spanning the −157/−14 region of the ICAM-3 promoter using nuclear extracts from THP-1, K-562 and Jurkat cells. The position of the major retarded species is indicated. B. EMSA was performed on the ICAM3.3 and ICAM3.5 oligonucleotides using nuclear extracts from the indicated COS-7 cells transfected with an empty expression vector (pCDNA3) or with either RUNX1 or RUNX3 together with CBF-β expression vector. The position of the RUNX1- and RUNX3-containing complex is shown. C. EMSA was performed on the ICAM3.5 and ICAM3.3 oligonucleotides using nuclear extracts from Jurkat cells in the absence (−) or presence of unlabeled competitor oligonucleotides (ICAM3.5, ICAM3.5mutRUNX, ICAM3.3, ICAM3.3mutRUNX, AMLcons) or polyclonal antisera against CD209 (Control antibody, Cnt Ab) or RUNX1 proteins (R-3034). The position of RUNX1-containing complexes are shown. Unlabeled competitor oligonucleotides were added at a 100-fold molar excess. D. EMSA was performed on the ICAM3.4 oligonucleotide using nuclear extracts from THP-1 cells in the absence (−) or presence of unlabeled competitor oligonucleotides (ICAM3.4, ICAM3.4mutCEBP, C/EBPcons) or polyclonal antibody against CD209 (Control antibody, Cnt Ab) or C/EBPα proteins (α-C/EBPα). The position of C/EBPα-containing complexes are shown. Unlabeled competitor oligonucleotides were added at a 100-fold molar excess. In A–D, EMSA's were performed twice with similar result and a representative experiment is shown. E. ICAM-3 promoter-based oligonucleotides with mutated nucleotides in lowercase and their relative positions. F. In vivo occupancy of the ICAM-3 promoter by RUNX1. Chromatin immunoprecipitation on Jurkat cells was performed with an affinity-purified polyclonal antisera specific for RUNX1 or purified rabbit IgG. Immunoprecipitated chromatin was analyzed by PCR using a pair of ICAM-3 promoter-specific primers that amplify a 234-bp fragment flanking the RUNX-binding sites at −80 and −29. ChIP experiment was performed twice with similar results, and a representative experiment is shown.
Techniques Used: Transfection, Expressing, Plasmid Preparation, In Vivo, Chromatin Immunoprecipitation, Affinity Purification, Purification, Immunoprecipitation, Binding Assay

Figure Legend Snippet: A. Disruption of the RUNX-binding elements leads to increased ICAM-3 gene promoter activity in THP-1 cells. THP-1 cells were transfected with the indicated reporter plasmids and luciferase activity was determined after 24 h. Promoter activity is expressed relative to the activity produced by the wild-type CD50-200Luc, arbitrarily set to 1, after normalization for transfection efficiency. Data represent mean ± SD of 6 independent experiments using two different DNA preparations. B. Knockdown on RUNX3 results in increased ICAM-3 mRNA levels. THP-1 cells were nucleofected with either siRNA for RUNX3 (two different RUNX3 specific-siRNA, siRNA RUNX3#1 and siRNA RUNX3#2) or a control siRNA (siRNA CNT). After 24 hours, total RNA was isolated and ICAM-3 mRNA was measured via quantitative RT-PCR (left). Results are expressed as Relative mRNA levels (relative to GAPDH mRNA levels and the ICAM-3 mRNA level in control siRNA-nucleofected cells). Data represent mean ± SD of 3 independent experiments. To confirm siRNA efficiency, one-fifth of the cells were lysed and underwent western blotting (right). The western blot was performed twice with similar results and one of the experiments is shown.
Techniques Used: Binding Assay, Activity Assay, Transfection, Luciferase, Produced, Isolation, Quantitative RT-PCR, Western Blot
thp1 cells (ATCC)


Structured Review

Thp1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/thp1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "In Vivo Inflammation Does Not Impair ABCA1-Mediated Cholesterol Efflux Capacity of HDL"
Article Title: In Vivo Inflammation Does Not Impair ABCA1-Mediated Cholesterol Efflux Capacity of HDL
Journal: Cholesterol
doi: 10.1155/2012/610741

Figure Legend Snippet: Cholesterol efflux capacity of MPO-HDL in fibroblasts and TP1 cells. HDL and MPO-HDL are used as cholesterol acceptors. Cholesterol efflux potential is measured in fibroblasts (white bars) and THP1 cells (black bars) under cholesterol-loaded conditions as described. The cells are labelled for 24 h, with 3 H-cholesterol in the absence (a) or presence (b) of LXR agonist. Efflux to HDL (10 μ g /mL apoA-I) is measured during a 4 h time span. Efflux medium without HDL is used as a negative control. Fractional cholesterol efflux is determined by measuring the release of radio-labeled cholesterol into the medium. The percentage efflux equals the counts in medium divided by total counts in medium and cells combined, subtracted by efflux to the control. Data are presented as mean ± SD; ** P < 0.01. Experiments were performed in triplicate.
Techniques Used: Negative Control, Labeling

Figure Legend Snippet: Efflux potential of HDL isolated from patients with Crohn's disease in fibroblasts and THP1 cells. Cholesterol acceptor capacity of UC-HDL (black bars) and PEG HDL (white bars) isolated from controls, and patients with active Crohn's disease and Crohn's disease in remission using cholesterol-laden fibroblasts and cholesterol-laden THP1 cells. Cells are labelled for 24 h with 3 H-cholesterol. Fibroblasts were incubated in the presence of LXR agonist TO901317. Efflux to HDL (10 μ g/mL apoA-I) is measured during a 4 h time span. Efflux medium without HDL is used as a negative control. Fractional cholesterol efflux is determined by measuring the release of radio-labeled cholesterol into the medium. The percentage efflux equals the counts in medium divided by total counts in medium and cells combined and was corrected for counts to medium without acceptor HDL. Experiments were performed in triplicate. Data are presented as mean ± SD; * P < 0.05.
Techniques Used: Isolation, Incubation, Negative Control, Labeling

Figure Legend Snippet: Efflux potential of UC-HDL and PEG-HDL isolated from patients with sepsis in fibroblasts and THP1 cells. Cholesterol acceptor capacity of UC-HDL (black bars) and PEG-HDL (white bars) from patients with sepsis using cholesterol-laden fibroblasts (and cholesterol-laden THP1 cells). Fibroblasts were incubated in the presence of LXR agonist TO901317. Cells are labelled for 24 h with 3 H-cholesterol. Efflux to HDL (10 μ g/mL apoA-I) is measured during a 4 h time span. Efflux medium without HDL is used as a negative control. Fractional cholesterol efflux is determined by measuring the release of radio-labeled cholesterol into the medium. The percentage efflux equals the counts in medium divided by total counts in medium and cells combined and was corrected for counts to the medium without acceptor HDL. Experiments were performed in triplicate. Data are presented as mean ± SD; * P < 0.05, ** P < 0.01.
Techniques Used: Isolation, Incubation, Negative Control, Labeling
human thp 1 cell lines (ATCC)


Structured Review

Human Thp 1 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human thp 1 cell lines/product/ATCC
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Interferon-Tau Attenuates Uptake of Nanoparticles and Secretion of Interleukin-1β in Macrophages"
Article Title: Interferon-Tau Attenuates Uptake of Nanoparticles and Secretion of Interleukin-1β in Macrophages
Journal: PLoS ONE
doi: 10.1371/journal.pone.0113974

Figure Legend Snippet: (A) THP-1 macrophages were incubated for 48 h with or without rbIFNT at the indicated concentrations. Subsequently, total RNA was extracted and analyzed by real-time RT-PCR for expression of ISG15 mRNA. (B–D) THP-1 macrophages were incubated for 48 h with rbIFNT or rhIFNB at the indicated concentrations. After priming with LPS (100 ng/mL) for 3 h or Pam3CSK4 (300 ng/ml) for 10 h, cells were treated with nano-silica particles (100 µg/mL) for 6 h. IL-1β (B and D) and IL-1α (C) levels in supernatants were then determined using ELISA. (E) LDH release from THP-1 macrophages treated with rbIFNT or rhIFNB. Data are expressed as means ±SEM (n = 3); Significant differences were identified using ANOVA; * p <0.05 and ** p <0.01.
Techniques Used: Incubation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Figure Legend Snippet: (A) THP-1 macrophages were treated with Green nano-silica for 6 h at the indicated concentrations. (B) THP-1 macrophages were incubated for 48 h with or without rbIFNT (2.7 and 27 IU/mL), and were then treated with Green nano-silica particles (30 µg/mL) for 6 h. After washing cells, uptake of Green nano-silica particles was analyzed by flow cytometry. (C) THP-1 macrophages were incubated for 1 h with or without cytochalasin D and were treated with Green nano-silica particles (30 µg/mL) for 6 h. After washing cells, uptake of Green nano-silica particles was analyzed by flow cytometry. Representative flow cytometry plots are presented. (D) THP-1 macrophages were incubated for 48 h with or without rbIFNT (27 IU/mL), and were then treated with nano-silica particles (30 µg/mL) for 6 h. Cells were stained with fluorescent-conjugated phalloidin and analyzed by flow cytometry. Nuclei were co-stained with Hoechst 33342. Representative confocal microscopic images of the F-actin assembly are presented. Quantitative analyses were performed and data are expressed as means ±SEM (n = 3); Significant differences were identified using ANOVA; * p <0.05 and ** p <0.01.
Techniques Used: Incubation, Flow Cytometry, Staining

Figure Legend Snippet: (A) THP-1 macrophages were incubated for 48 h with or without rbIFNT (27 IU/mL) and total RNA was then extracted and analyzed by real-time RT-PCR for MARCO, CD36, SR-BI, MSR1 and OLR1 mRNA. (B) THP-1 macrophages were transfected with MARCO siRNA (siMARCO) or control siRNA (siN.C.) and analyzed MARCO mRNA expression by real-time RT-PCR (C). THP-1 macrophages transfected with MARCO siRNA or control siRNA were incubated with green nano-silica particles (30 µg/mL) for 6 h. After washing cells, uptake of green nano-silica particles was analyzed the amount of nano-silica (mean fluorescence intensity; MFI) by flow cytometry. After nano-silica treatment, IL-1β levels in supernatants were determined using ELISA. Data are expressed as means ±SEM (n = 3–4); Significant differences were identified using t-test; ** p <0.01 vs. control.
Techniques Used: Incubation, Quantitative RT-PCR, Transfection, Expressing, Fluorescence, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Figure Legend Snippet: (A) THP-1 macrophages were treated with nano-silica (100 µg/mL) for the indicated periods. ROS generation was assessed using DCFDA and flow cytometry. (B) THP-1 macrophages were incubated for 48 h with or without rbIFNT (27 U/mL) and were treated with nano-silica particles (100 µg/mL) for 6 h. ROS generation was assessed using DCFDA by flow cytometry. Quantitative analyses were performed and data are expressed as means ±SEM (n = 3); Significant differences were identified using ANOVA; * p <0.05 and ** p <0.01. Representative confocal microscopic images of ROS production in THP-1 macrophages; Nuclei were co-stained with Hoechst 33342.
Techniques Used: Flow Cytometry, Incubation, Staining

Figure Legend Snippet: (A) THP-1 macrophages were incubated for 48 h with or without rbIFNT. After priming with LPS (100 ng/mL) for 3 h, cells were treated with nano-silica particles (100 µg/mL) for 6 h. Protein levels of pro-IL-1β and mature IL-1β in supernatants were detected by Western blot analyses. Representative photographs are shown. (B) Subsequently, total RNA was extracted and analyzed by real-time RT-PCR for expression of IL-1β mRNA. (C) IL-10 levels in supernatants were then determined using ELISA. (D) THP-1 macrophages were incubated with or without rbIFNT (27 IU/mL) in the presence of antibody against human IL-10 or control IgG, and then treated with LPS and nano-silica. IL-1β levels in supernatants were then determined using ELISA. Data are expressed as means ±SEM (n = 3); Significant differences were identified using ANOVA; ** p <0.01.
Techniques Used: Incubation, Western Blot, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay
kb 3 1 cells (ATCC)


Structured Review
Kb 3 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kb 3 1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
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thp1 cells (ATCC)


Structured Review

Thp1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/thp1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Analysis of MicroRNA-Mediated Translation Activation of In Vitro Transcribed Reporters in Quiescent Cells"
Article Title: Analysis of MicroRNA-Mediated Translation Activation of In Vitro Transcribed Reporters in Quiescent Cells
Journal: Methods in molecular biology (Clifton, N.J.)
doi: 10.1007/978-1-4939-7371-2_18

Figure Legend Snippet: 100 ng of in vitro transcribed capped CX-Firefly luciferase mRNA—without a poly(A) tail (CXA0) and with cordycepin added at the 3′ end—was nucleofected along with 20 ng of renilla plasmid, and 500 pmol of miRcxcr4 or control miR30a, into 1 × 106 THP1 cells. After nucleofection, cells were grown in RPMI medium supplemented with 10% fetal bovine serum (FBS) for 24 h, and then shifted to G0 medium (RPMI medium without FBS) for 42 h before analysis of luciferase activity. More than threefold increase in the translation of CX-firefly luciferase is observed in the presence of cognate microRNA miRcxcr4 compared to control microRNA miR30a
Techniques Used: In Vitro, Luciferase, Plasmid Preparation, Activity Assay

Figure Legend Snippet: 1 × 106 THP1 cells were nucleofected with 2.0 μg of pmax-GFP, using Kit V and nucleofector device II from Lonza. Left and right panels show 20× phase-contrast image and green fluorescent protein (GFP) positive THP1 cells respectively (images captured using a Leica DMI-4000B fluorescence inverted microscope). (A) Nucleofection program U-001 for high cell viability was used and transfection efficiency of about 24% was observed. (B) Program V-001 for high nucleofection efficiency was used and transfection efficiency of about 40% was achieved
Techniques Used: Fluorescence, Inverted Microscopy, Transfection
human monocytic cell line thp1 cells (ATCC)


Structured Review

Human Monocytic Cell Line Thp1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human monocytic cell line thp1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Mycobacterium tuberculosis infection elevates SLIT2 expression to modulate oxidative stress responses in macrophages"
Article Title: Mycobacterium tuberculosis infection elevates SLIT2 expression to modulate oxidative stress responses in macrophages
Journal: bioRxiv
doi: 10.1101/2022.10.13.512188

Figure Legend Snippet: (A) THP1 macrophages were infected with Mtb H37Rv for the indicated time points. Whole cell lysates were assessed for SLIT2 protein expression by immunoblotting. (B) Mouse peritoneal macrophages were infected with Mtb H37Rv for 24 h and assessed for the transcript levels of ROBO receptors. (C) Mouse peritoneal macrophages were infected with Mtb H37Rv for the indicated time points and ROBO2 levels were assessed by immunoblotting. All immunoblotting data are representative of three independent experiments. β-ACTIN was utilized as loading control. qRT-PCR data represents mean±S.E.M. from three independent experiments.
Techniques Used: Infection, Expressing, Western Blot, Quantitative RT-PCR

Figure Legend Snippet: (A) Schematic representing the probable mechanism leading to Slit2 upregulation during Mtb infection. (B) THP1 macrophages were infected with Mtb H37Rv for 24 h and assessed for the recruitment of H3K27me3 and p-Histone H3 (Ser28) over the Slit2 promoter by ChIP assay. (C) Mouse peritoneal macrophages were treated with the indicated inhibitors for 1 h, followed by 24 h infection with Mtb H37Rv and assessed for the expression of Slit ligands transcripts by qRT-PCR (D) Mouse peritoneal macrophages were treated with the indicated inhibitors for 1 h, followed by 24 h infection with Mtb H37Rv and assessed for the expression of indicated proteins by immunoblotting (E, F) Mouse peritoneal macrophages were treated with the indicated inhibitors for 1 h. Cells were infected with Mtb H37Rv for 24 h and assessed for the accumulation of ROS by fluorescence microscopy (CellROX Deep Red); (E) representative image and (F) respective quantification. . All immunoblotting and immunofluorescence data are representative of three independent experiments. β-ACTIN was utilized as loading control. qRT-PCR data represents mean±S.E.M. from three independent experiments. ROS, reactive oxygen species; h, hours; ChIP, Chromatin immunoprecipitation; d, days; kda, kilodalton; CTCF, corrected total cell fluorescence. *, p<0.05; ****, p < 0.0001 (Student’s t-test in B, One-way ANOVA in C,F; GraphPad Prism 6.0 and 9.0). Scale bar, 5 μm. SB203580: p38 inhibitor, SP600125: JNK1/2 inhibitor.
Techniques Used: Infection, Expressing, Quantitative RT-PCR, Western Blot, Fluorescence, Microscopy, Immunofluorescence, Chromatin Immunoprecipitation

Figure Legend Snippet: (A) THP1 macrophages were infected with Mtb H37Rv for 24 h and assessed for the recruitment of H3K27me3 over the Slit2 promoter by ChIP assay. (B) Mouse peritoneal macrophages were infected with Mtb H37Rv for the indicated time points and assessed for the levels of p-Histone H3 (Ser28) by immunoblotting. (C) THP1 macrophages were infected with Mtb H37Rv for the indicated time points and assessed for the levels of p-Histone H3 (Ser28) by immunoblotting. (D) THP1 macrophages were treated with the indicated inhibitors for 1 h, followed by 24 h infection with Mtb H37Rv and assessed for the expression of SLIT2 by immunoblotting. All immunoblotting data are representative of three independent experiments. β-ACTIN was utilized as loading control.
Techniques Used: Infection, Western Blot, Expressing

Figure Legend Snippet: (A) THP1 macrophages were infected with Mtb H37Rv for 24 h and assessed for the levels of VNN1 by immunoblotting. (B) Mouse peritoneal macrophages were treated with rSLIT2 at the indicated concentrations for 12 h and assessed for the levels of VNN1 by immunoblotting. (C) Mouse peritoneal macrophages were transfected with NT or Vnn1 siRNAs. Transfected cells were infected with Mtb H37Rv for 24 h and assessed for the accumulation of ROS by fluorescence microscopy (CellROX Deep Red). All immunoblotting and immunofluorescence data are representative of three independent experiments. β-ACTIN was utilized as loading control. Scale bar, 5 μm.
Techniques Used: Infection, Western Blot, Transfection, Fluorescence, Microscopy, Immunofluorescence
thp1 cells (ATCC)


Structured Review
Thp1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/thp1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
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monocytic cells (ATCC)


Structured Review
Monocytic Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/monocytic cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
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kb prophage 1 (ATCC)


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Kb Prophage 1, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/kb prophage 1/product/ATCC
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Comparative genomics of Acinetobacter baumannii and therapeutic bacteriophages from a patient undergoing phage therapy"
Article Title: Comparative genomics of Acinetobacter baumannii and therapeutic bacteriophages from a patient undergoing phage therapy
Journal: Nature Communications
doi: 10.1038/s41467-022-31455-5

Figure Legend Snippet: A DNA sequence relatedness of six T4-like myophages, showing pairwise percent DNA sequence identities as determined by ProgressiveMauve (upper section) and DNA dotplots visually representing DNA sequence alignments between phages (lower section). B Protein sequence-based relationships of 2834 Caudoviricetes phages representing all species in the ICTV taxonomy, plus the seven treatment phages and two prophages identified in strains TP1, TP2, and TP3. Unclustered singletons (17 in total) are removed from the visualization. Distinct colors are assigned at the Subfamily level; if no Subfamily was assigned, color is assigned at the Family level. Circled clusters are enlarged in C – E as labeled. C Enlarged cluster representing the Autographiviridae . Nodes are colored based on their Subfamily membership, with the legend identifying prominent clades; the node representing phage AbTP3phi1 is outlined in black, and nodes representing clade-founding phages T7, phiKMV and Fri1 are labeled. D Enlarged cluster containing the T4-like subfamilies, including the Twarogvirinae ; this large cluster is also linked to the T5-like Markadamsvirinae and clusters of diverse myophages including the V5-like Vequintaviridae and FelixO1-like Ounavirinae . Nodes are colored based on their Subfamily membership with the legend identifying prominent clades. Nodes representing the six treatment myophages are outlined in black, and nodes representing clade-founding phages T4, T5, FelixO1 and V5 are labeled. E Enlarged cluster containing the two prophage elements identified in strains TP1, TP2 and TP3. These prophages are not closely related to other classified phages, with the 52 kb prophage 1 distantly linked to the Guernseyvirinae , and the 42 kb prophage 2 related to two other unclassified siphophages.
Techniques Used: Sequencing, Labeling
thp1 cells (ATCC)


Structured Review

Thp1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/thp1 cells/product/ATCC
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Self-guarding of MORC3 enables virulence factor-triggered immunity"
Article Title: Self-guarding of MORC3 enables virulence factor-triggered immunity
Journal: Nature
doi: 10.1038/s41586-021-04054-5

Figure Legend Snippet: PMA-differentiated THP1 (a, c) or U937 (b) human myeloid-like cells were infected with HSV-1 at MOI=1 or indicated MOI for 24h (a, b) or indicated timepoints (c) or transfected with 2ʹ-3ʹ-cGAMP for 4h. Gene-expression analysis is depicted as mean + SEM of three independent experiments. ND = not detected. * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significantly different than WT, tested by two-way ANOVA or Mixed-effects model and Bonferroni’s post hoc test. See Source Data for exact p values.
Techniques Used: Infection, Transfection, Expressing

Figure Legend Snippet: BLaER1 monocytes (a), PMA-differentiated THP1 (b) or U937 (c) cells expressing doxycycline-inducible virulence factors were stimulated with doxycycline for 24h, left untreated or stimulated with DNA for 3h. Gene-expression as quantified by q-RT-PCR is depicted as mean + SEM of three independent experiments. IFNB1 expression levels in a) are partially duplicated from Fig. 1c. * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significantly different than the corresponding mCherry-expressing condition, tested by two-way ANOVA or Mixed-effects model and Dunnett’s post hoc test. ND = not detected. See Source Data for exact p values.
Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

Figure Legend Snippet: (a-d) Gene-expression in PMA-differentiated THP1-Cas9 or PMA-differentiated U937-Cas9 human myeloid-like cells expressing indicated sgRNAs is depicted as mean + SEM of three independent experiments. Protein expression in the same cells was analyzed by immunoblot. ND = not detected. *** p < 0.001; ns = not significantly different than the corresponding scramble sgRNA-expressing condition, tested by two-way ANOVA and Bonferroni’s post hoc test. See Source Data for exact p values.
Techniques Used: Expressing, Western Blot