human monocytic cell line u937  (ATCC)


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    ATCC human monocytic cell line u937
    L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human <t>U937</t> macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant
    Human Monocytic Cell Line U937, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Effects of Leea indica leaf extracts and its phytoconstituents on natural killer cell-mediated cytotoxicity in human ovarian cancer"

    Article Title: Effects of Leea indica leaf extracts and its phytoconstituents on natural killer cell-mediated cytotoxicity in human ovarian cancer

    Journal: BMC Complementary Medicine and Therapies

    doi: 10.1186/s12906-023-03904-1

    L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human U937 macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant
    Figure Legend Snippet: L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human U937 macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control

    human monocyte cell line thp 1 cells  (ATCC)


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    ATCC human monocyte cell line thp 1 cells
    Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced <t>THP-1</t> cells. *** P < 0.001 vs. Control group
    Human Monocyte Cell Line Thp 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "MicroRNA-218-5p regulates inflammation response via targeting TLR4 in atherosclerosis"

    Article Title: MicroRNA-218-5p regulates inflammation response via targeting TLR4 in atherosclerosis

    Journal: BMC Cardiovascular Disorders

    doi: 10.1186/s12872-023-03124-y

    Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced THP-1 cells. *** P < 0.001 vs. Control group
    Figure Legend Snippet: Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced THP-1 cells. *** P < 0.001 vs. Control group

    Techniques Used: Binding Assay, Luciferase, Reporter Gene Assay, Expressing

    murine monocytic cell line raw264 7  (ATCC)


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    ATCC murine monocytic cell line raw264 7
    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor <t>(RAW264.7)</t> cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Murine Monocytic Cell Line Raw264 7, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein"

    Article Title: Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein

    Journal: Journal of Extracellular Vesicles

    doi: 10.1002/jev2.12312

    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor (RAW264.7) cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Figure Legend Snippet: Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor (RAW264.7) cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.

    Techniques Used: Derivative Assay, Tomography, Cell Culture, Staining, Light Microscopy, Activity Assay, Fluorescence

    RNA‐seq confirmed osteoclastogenesis induced by prostate cancer‐derived EVs. (a) Schematic protocol for the gene expression analysis in osteoclast precursor cells (RAW264.7 cells) that were treated with EVs derived from PC3M cells (30 μg/mL) and RANKL (10 ng/mL). (b) Heatmap showing the difference in the expression level of mRNA that was previously reported to be related to osteoclastogenesis between osteoclast precursor cells treated with EVs derived from PC3M cells and RANKL. (c) The difference in NFATc1, TRAP, DC‐STAMP and CTSK expression between the samples. The y‐axis represents the expression level of the transcript quantified as reads per kilobase of the transcript per million mapped reads to the transcriptome (RPKM). (d) Osteoclast precursor cells were cultured with EVs derived from PC3M cells (30 μg/mL) with or without RANKL (10 ng/mL). After 96 h incubation, quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK. (e) Quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK after 168 h incubation. Error bars represent s.d. of triplicate experiments. Corrections passed the Bonferroni threshold ( p < 0.05/3 = 0.0167) are marked with asterisk (*). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs and EV30 represents 30 μg/mL EVs.
    Figure Legend Snippet: RNA‐seq confirmed osteoclastogenesis induced by prostate cancer‐derived EVs. (a) Schematic protocol for the gene expression analysis in osteoclast precursor cells (RAW264.7 cells) that were treated with EVs derived from PC3M cells (30 μg/mL) and RANKL (10 ng/mL). (b) Heatmap showing the difference in the expression level of mRNA that was previously reported to be related to osteoclastogenesis between osteoclast precursor cells treated with EVs derived from PC3M cells and RANKL. (c) The difference in NFATc1, TRAP, DC‐STAMP and CTSK expression between the samples. The y‐axis represents the expression level of the transcript quantified as reads per kilobase of the transcript per million mapped reads to the transcriptome (RPKM). (d) Osteoclast precursor cells were cultured with EVs derived from PC3M cells (30 μg/mL) with or without RANKL (10 ng/mL). After 96 h incubation, quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK. (e) Quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK after 168 h incubation. Error bars represent s.d. of triplicate experiments. Corrections passed the Bonferroni threshold ( p < 0.05/3 = 0.0167) are marked with asterisk (*). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs and EV30 represents 30 μg/mL EVs.

    Techniques Used: RNA Sequencing Assay, Derivative Assay, Expressing, Cell Culture, Incubation, Real-time Polymerase Chain Reaction

    Prostate cancer‐derived EVs specifically promote osteoclastogenesis. (a) Representative images of EVs isolated from PC3M and PNT2 cells and observed under transmission electron microscopy. Scale bar, 100 nm. (b) The particle size distributions and concentrations of EVs from PC3M and PNT2 cells were measured using a NanoSight nanoparticle tracking system. (c) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐CD9, anti‐CD63, and anti‐CD81 antibodies. (d) Representative confocal microscopy images. EVs derived from PC3M and PNT2 cells were labelled with PKH67 and added to osteoclast precursors in the presence of RANKL (10 ng/mL). White dots indicate cell morphologies. Scale bars, 25 μm. (e) Analysis of the effect of EVs derived from PC3M and PNT2 cells on osteoclast precursors (RAW264.7 cells). EVs were added to RAW264.7 cells, and osteoclast differentiation was evaluated by tartrate‐resistant acid phosphatase (TRAP) staining after 96 h. The number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (f) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐RANKL and anti‐NFATc1 antibodies. As positive controls, recombinant RANKL and cell lysates from osteoclast cells that were differentiated from RAW264.7 cells were used. R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL EVs and EV30 represents 30 μg/mL EVs. Error bars represent s.d. of triplicate experiments, with significant differences determined by Dunnett's test (** p < 0.01)
    Figure Legend Snippet: Prostate cancer‐derived EVs specifically promote osteoclastogenesis. (a) Representative images of EVs isolated from PC3M and PNT2 cells and observed under transmission electron microscopy. Scale bar, 100 nm. (b) The particle size distributions and concentrations of EVs from PC3M and PNT2 cells were measured using a NanoSight nanoparticle tracking system. (c) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐CD9, anti‐CD63, and anti‐CD81 antibodies. (d) Representative confocal microscopy images. EVs derived from PC3M and PNT2 cells were labelled with PKH67 and added to osteoclast precursors in the presence of RANKL (10 ng/mL). White dots indicate cell morphologies. Scale bars, 25 μm. (e) Analysis of the effect of EVs derived from PC3M and PNT2 cells on osteoclast precursors (RAW264.7 cells). EVs were added to RAW264.7 cells, and osteoclast differentiation was evaluated by tartrate‐resistant acid phosphatase (TRAP) staining after 96 h. The number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (f) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐RANKL and anti‐NFATc1 antibodies. As positive controls, recombinant RANKL and cell lysates from osteoclast cells that were differentiated from RAW264.7 cells were used. R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL EVs and EV30 represents 30 μg/mL EVs. Error bars represent s.d. of triplicate experiments, with significant differences determined by Dunnett's test (** p < 0.01)

    Techniques Used: Derivative Assay, Isolation, Transmission Assay, Electron Microscopy, Western Blot, Confocal Microscopy, Staining, Recombinant

    human monocytic leukemia cells  (ATCC)


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    ATCC human monocytic leukemia cells
    Human Monocytic Leukemia Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human monocytic leukemia thp 1 cells  (ATCC)


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    ATCC human monocytic leukemia thp 1 cells
    Human Monocytic Leukemia Thp 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human acute monocyte leukemia cell line  (ATCC)


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    ATCC human acute monocyte leukemia cell line
    Human Acute Monocyte Leukemia Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human monocytic leukemia thp 1 cells  (ATCC)


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    ATCC human monocytic leukemia thp 1 cells
    Human Monocytic Leukemia Thp 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    monocytic leukemia cell line thp  (ATCC)


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    ATCC monocytic leukemia cell line thp
    SB reduced the expression of inflammatory genes in stimulated <t>THP-1</t> cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.
    Monocytic Leukemia Cell Line Thp, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Sodium benzoate attenuates 2,8-dihydroxyadenine nephropathy by inhibiting monocyte/macrophage TNF-α expression"

    Article Title: Sodium benzoate attenuates 2,8-dihydroxyadenine nephropathy by inhibiting monocyte/macrophage TNF-α expression

    Journal: Scientific Reports

    doi: 10.1038/s41598-023-30056-6

    SB reduced the expression of inflammatory genes in stimulated THP-1 cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.
    Figure Legend Snippet: SB reduced the expression of inflammatory genes in stimulated THP-1 cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transduction, Software

    Transwell assay shows that SB reduces THP-1 migration. ( a ) The total number of cells in each well in the lower chamber of the 5 μm-pore transwell were counted and plotted after 24 h of incubation (n = 6 each). Cell counts per well were significantly lower in the FBS + SB group compared with the FBS group. ( b ) SB dose dependency was confirmed by applying different concentrations of SB to the upper chamber of the transwell (n = 4–6). Higher SB concentrations significantly reduced the number of migrating cells. * p < 0.05; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. FBS, fetal bovine serum; SB, sodium benzoate.
    Figure Legend Snippet: Transwell assay shows that SB reduces THP-1 migration. ( a ) The total number of cells in each well in the lower chamber of the 5 μm-pore transwell were counted and plotted after 24 h of incubation (n = 6 each). Cell counts per well were significantly lower in the FBS + SB group compared with the FBS group. ( b ) SB dose dependency was confirmed by applying different concentrations of SB to the upper chamber of the transwell (n = 4–6). Higher SB concentrations significantly reduced the number of migrating cells. * p < 0.05; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. FBS, fetal bovine serum; SB, sodium benzoate.

    Techniques Used: Transwell Assay, Migration, Incubation

    SB reduces NF-κB RelB levels of protein, mRNA, and binding to motif DNA sequence. ( a ) Band intensities of NF-κB RelB relative to beta-actin were plotted. In AdCKD kidney, NF-κB RelB protein expression was increased compared to control kidney, which was significantly decreased in AdCKD + SB kidney. ( b ) NF-κB RelB protein expression was increased in LPS stimulated THP-1 cells compared to control cells, which was significantly decreased in SB treated LPS stimulated THP-1 cells. ( c ) mRNA levels of NF-κB RelB in AdCKD kidney were increased compared to control, which was significantly decreased in AdCKD + SB. ( d ) mRNA levels of NF-κB RelB in LPS stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( e ) mRNA levels of NF-κB RelB in TNF-α stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated TNF-α stimulated cells. ( f ) NF-κB RelB binding to motif DNA sequence was increased in AdCKD kidney compared to control, which was significantly decreased in AdCKD + SB kidney. ( g ) NF-κB RelB binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( h ) NF-κB p65 binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was similar levels compared to SB-treated LPS stimulated cells. ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; SB, sodium benzoate.
    Figure Legend Snippet: SB reduces NF-κB RelB levels of protein, mRNA, and binding to motif DNA sequence. ( a ) Band intensities of NF-κB RelB relative to beta-actin were plotted. In AdCKD kidney, NF-κB RelB protein expression was increased compared to control kidney, which was significantly decreased in AdCKD + SB kidney. ( b ) NF-κB RelB protein expression was increased in LPS stimulated THP-1 cells compared to control cells, which was significantly decreased in SB treated LPS stimulated THP-1 cells. ( c ) mRNA levels of NF-κB RelB in AdCKD kidney were increased compared to control, which was significantly decreased in AdCKD + SB. ( d ) mRNA levels of NF-κB RelB in LPS stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( e ) mRNA levels of NF-κB RelB in TNF-α stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated TNF-α stimulated cells. ( f ) NF-κB RelB binding to motif DNA sequence was increased in AdCKD kidney compared to control, which was significantly decreased in AdCKD + SB kidney. ( g ) NF-κB RelB binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( h ) NF-κB p65 binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was similar levels compared to SB-treated LPS stimulated cells. ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; SB, sodium benzoate.

    Techniques Used: Binding Assay, Sequencing, Expressing

    murine macrophage monocyte cells  (ATCC)


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    ATCC murine macrophage monocyte cells
    Murine Macrophage Monocyte Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    murine macrophage monocyte cells  (ATCC)


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    Structured Review

    ATCC murine macrophage monocyte cells
    Murine Macrophage Monocyte Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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  • 86
    ATCC human monocytic cell line u937
    L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human <t>U937</t> macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant
    Human Monocytic Cell Line U937, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC human monocyte cell line thp 1 cells
    Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced <t>THP-1</t> cells. *** P < 0.001 vs. Control group
    Human Monocyte Cell Line Thp 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC murine monocytic cell line raw264 7
    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor <t>(RAW264.7)</t> cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Murine Monocytic Cell Line Raw264 7, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC human monocytic leukemia cells
    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor <t>(RAW264.7)</t> cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Human Monocytic Leukemia Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC human monocytic leukemia thp 1 cells
    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor <t>(RAW264.7)</t> cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Human Monocytic Leukemia Thp 1 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC human acute monocyte leukemia cell line
    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor <t>(RAW264.7)</t> cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.
    Human Acute Monocyte Leukemia Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC monocytic leukemia cell line thp
    SB reduced the expression of inflammatory genes in stimulated <t>THP-1</t> cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.
    Monocytic Leukemia Cell Line Thp, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC murine macrophage monocyte cells
    SB reduced the expression of inflammatory genes in stimulated <t>THP-1</t> cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.
    Murine Macrophage Monocyte Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human U937 macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant

    Journal: BMC Complementary Medicine and Therapies

    Article Title: Effects of Leea indica leaf extracts and its phytoconstituents on natural killer cell-mediated cytotoxicity in human ovarian cancer

    doi: 10.1186/s12906-023-03904-1

    Figure Lengend Snippet: L. indica crude leaf extract, ethyl acetate fraction, and methyl gallate suppressed TNF-α and IL-1β production by human U937 macrophages. Fold change of TNF-α ( A , B ) and IL-1β ( C , D ) production relative to control in the supernatant of human U937 macrophages measured by ELISA. Cells were pretreated for 5 h with or without 40 μg/mL L. indica crude leaf extract, 10 μg/mL L. indica ethyl acetate fraction (EA), 38 μM methyl gallate (MG) or 38 μM gallic acid (GA), followed by LPS stimulation. Cytokine production by cells treated with both PMA and LPS was taken as 1. Dexamethasone (DEX) at 64.4 ng/mL was used as positive control, while DMSO was used as negative control. Results are presented as mean fold change ± SD of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant

    Article Snippet: Human advanced ovarian cancer cell lines OVCAR-5 (NCI Frederick, USA) and SK-OV-3 (ATCC, HTB-77), human monocytic cell line U937 (ATCC, CRL-1593.2), human NK cell line NK-92 (ATCC, CRL-2407) were purchased.

    Techniques: Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control

    Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced THP-1 cells. *** P < 0.001 vs. Control group

    Journal: BMC Cardiovascular Disorders

    Article Title: MicroRNA-218-5p regulates inflammation response via targeting TLR4 in atherosclerosis

    doi: 10.1186/s12872-023-03124-y

    Figure Lengend Snippet: Prediction and verification of miR-218-5p target genes. (A) . Complementary binding sites of miR-218-5p and TLR4. (B) . Luciferase reporter gene assay confirmed that miR-218-5p directly targets TLR4. (C) . TLR4 expression was increased in ox-LDL-induced THP-1 cells. *** P < 0.001 vs. Control group

    Article Snippet: The human monocyte cell line THP-1 cells were purchased from ATCC.

    Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Expressing

    Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor (RAW264.7) cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.

    Journal: Journal of Extracellular Vesicles

    Article Title: Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein

    doi: 10.1002/jev2.12312

    Figure Lengend Snippet: Prostate cancer‐derived EVs promote osteoclastogenesis. (a) Intratibial tumours were evaluated by the photon radiance of the cancer cell bioluminescence (left panel). Microcomputed tomography (μCT) revealed the same area as the osteolytic tumour site (yellow arrow in right panel). (b) Osteoclast precursor (RAW264.7) cells were cultured with EVs derived from PC3M cells with or without RANKL. At the end of Day 4, osteoclast precursor‐induced cells were fixed, stained for TRAP, and observed under a light microscope. Scale bar, 100 μm. The enlarged images are shown at the upper left of each image. (c) Osteoclast precursor cells were cultured with EVs derived from PC3M cells with or without RANKL. At different time points, osteoclast precursor‐induced cells were fixed and stained for TRAP, and the number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (d) The effect of PC3M‐derived EVs on bone resorption activity of osteoclasts was evaluated by fluorescence intensity. Error bars represent s.d. of triplicate experiments. * and ** indicate significant differences from the RANKL‐stimulated group (R10) as determined by Dunnett's test (* p < 0.05, ** p < 0.01). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs, and EV30 represents 30 μg/mL PC3M‐derived EVs.

    Article Snippet: The murine monocytic cell line RAW264.7 was purchased from American Type Culture Collection.

    Techniques: Derivative Assay, Tomography, Cell Culture, Staining, Light Microscopy, Activity Assay, Fluorescence

    RNA‐seq confirmed osteoclastogenesis induced by prostate cancer‐derived EVs. (a) Schematic protocol for the gene expression analysis in osteoclast precursor cells (RAW264.7 cells) that were treated with EVs derived from PC3M cells (30 μg/mL) and RANKL (10 ng/mL). (b) Heatmap showing the difference in the expression level of mRNA that was previously reported to be related to osteoclastogenesis between osteoclast precursor cells treated with EVs derived from PC3M cells and RANKL. (c) The difference in NFATc1, TRAP, DC‐STAMP and CTSK expression between the samples. The y‐axis represents the expression level of the transcript quantified as reads per kilobase of the transcript per million mapped reads to the transcriptome (RPKM). (d) Osteoclast precursor cells were cultured with EVs derived from PC3M cells (30 μg/mL) with or without RANKL (10 ng/mL). After 96 h incubation, quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK. (e) Quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK after 168 h incubation. Error bars represent s.d. of triplicate experiments. Corrections passed the Bonferroni threshold ( p < 0.05/3 = 0.0167) are marked with asterisk (*). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs and EV30 represents 30 μg/mL EVs.

    Journal: Journal of Extracellular Vesicles

    Article Title: Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein

    doi: 10.1002/jev2.12312

    Figure Lengend Snippet: RNA‐seq confirmed osteoclastogenesis induced by prostate cancer‐derived EVs. (a) Schematic protocol for the gene expression analysis in osteoclast precursor cells (RAW264.7 cells) that were treated with EVs derived from PC3M cells (30 μg/mL) and RANKL (10 ng/mL). (b) Heatmap showing the difference in the expression level of mRNA that was previously reported to be related to osteoclastogenesis between osteoclast precursor cells treated with EVs derived from PC3M cells and RANKL. (c) The difference in NFATc1, TRAP, DC‐STAMP and CTSK expression between the samples. The y‐axis represents the expression level of the transcript quantified as reads per kilobase of the transcript per million mapped reads to the transcriptome (RPKM). (d) Osteoclast precursor cells were cultured with EVs derived from PC3M cells (30 μg/mL) with or without RANKL (10 ng/mL). After 96 h incubation, quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK. (e) Quantitative PCR analysis was performed for markers of osteoclast differentiation, including TRAP, DC‐STAMP and CTSK after 168 h incubation. Error bars represent s.d. of triplicate experiments. Corrections passed the Bonferroni threshold ( p < 0.05/3 = 0.0167) are marked with asterisk (*). R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL PC3M‐derived EVs and EV30 represents 30 μg/mL EVs.

    Article Snippet: The murine monocytic cell line RAW264.7 was purchased from American Type Culture Collection.

    Techniques: RNA Sequencing Assay, Derivative Assay, Expressing, Cell Culture, Incubation, Real-time Polymerase Chain Reaction

    Prostate cancer‐derived EVs specifically promote osteoclastogenesis. (a) Representative images of EVs isolated from PC3M and PNT2 cells and observed under transmission electron microscopy. Scale bar, 100 nm. (b) The particle size distributions and concentrations of EVs from PC3M and PNT2 cells were measured using a NanoSight nanoparticle tracking system. (c) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐CD9, anti‐CD63, and anti‐CD81 antibodies. (d) Representative confocal microscopy images. EVs derived from PC3M and PNT2 cells were labelled with PKH67 and added to osteoclast precursors in the presence of RANKL (10 ng/mL). White dots indicate cell morphologies. Scale bars, 25 μm. (e) Analysis of the effect of EVs derived from PC3M and PNT2 cells on osteoclast precursors (RAW264.7 cells). EVs were added to RAW264.7 cells, and osteoclast differentiation was evaluated by tartrate‐resistant acid phosphatase (TRAP) staining after 96 h. The number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (f) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐RANKL and anti‐NFATc1 antibodies. As positive controls, recombinant RANKL and cell lysates from osteoclast cells that were differentiated from RAW264.7 cells were used. R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL EVs and EV30 represents 30 μg/mL EVs. Error bars represent s.d. of triplicate experiments, with significant differences determined by Dunnett's test (** p < 0.01)

    Journal: Journal of Extracellular Vesicles

    Article Title: Metastatic prostate cancer‐derived extracellular vesicles facilitate osteoclastogenesis by transferring the CDCP1 protein

    doi: 10.1002/jev2.12312

    Figure Lengend Snippet: Prostate cancer‐derived EVs specifically promote osteoclastogenesis. (a) Representative images of EVs isolated from PC3M and PNT2 cells and observed under transmission electron microscopy. Scale bar, 100 nm. (b) The particle size distributions and concentrations of EVs from PC3M and PNT2 cells were measured using a NanoSight nanoparticle tracking system. (c) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐CD9, anti‐CD63, and anti‐CD81 antibodies. (d) Representative confocal microscopy images. EVs derived from PC3M and PNT2 cells were labelled with PKH67 and added to osteoclast precursors in the presence of RANKL (10 ng/mL). White dots indicate cell morphologies. Scale bars, 25 μm. (e) Analysis of the effect of EVs derived from PC3M and PNT2 cells on osteoclast precursors (RAW264.7 cells). EVs were added to RAW264.7 cells, and osteoclast differentiation was evaluated by tartrate‐resistant acid phosphatase (TRAP) staining after 96 h. The number of TRAP‐positive multinucleated cells containing more than 3 nuclei (MNCs) was counted. (f) EVs from PC3M and PNT2 cells were analysed by immunoblotting using anti‐RANKL and anti‐NFATc1 antibodies. As positive controls, recombinant RANKL and cell lysates from osteoclast cells that were differentiated from RAW264.7 cells were used. R10 represents 10 ng/mL RANKL, EV10 represents 10 μg/mL EVs and EV30 represents 30 μg/mL EVs. Error bars represent s.d. of triplicate experiments, with significant differences determined by Dunnett's test (** p < 0.01)

    Article Snippet: The murine monocytic cell line RAW264.7 was purchased from American Type Culture Collection.

    Techniques: Derivative Assay, Isolation, Transmission Assay, Electron Microscopy, Western Blot, Confocal Microscopy, Staining, Recombinant

    SB reduced the expression of inflammatory genes in stimulated THP-1 cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.

    Journal: Scientific Reports

    Article Title: Sodium benzoate attenuates 2,8-dihydroxyadenine nephropathy by inhibiting monocyte/macrophage TNF-α expression

    doi: 10.1038/s41598-023-30056-6

    Figure Lengend Snippet: SB reduced the expression of inflammatory genes in stimulated THP-1 cells. ( a ) Protocol for LPS stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( b ) TNF-α , ( c ) IL-1β , and ( d ) MCP-1 . mRNA levels were adjusted to that of GAPDH (n = 5–10 each). ( e ) Protocol for TNF-α stimulation in THP-1 cells with or without SB preincubation. RT-PCR analysis of ( f ) TNF-α , ( g ) IL-1β , and ( h ) MCP-1. mRNA levels were adjusted to that of GAPDH (n = 6 each). ( i ) Immunoblotting (left panel) and quantification (right panel) of phosphorylated signal transduction molecules (n = 5–6). Band intensities were quantified using ImageJ software and adjusted to the intensity of the internal control molecule, β-actin. LPS (lane 2) significantly increased phospho-JNK and phospho-NF-κB p65 compared with the control (lane 1). SB did not decrease the phosphorylation of either molecule (lane 4). For comparison, the MEK inhibitor U0126 significantly decreased phospho-ERK levels (lane 5), while the JNK inhibitor SP600125 significantly decreased phospho-JNK (lane 6) and phospho-Akt (lane 6) levels. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; SB, sodium benzoate.

    Article Snippet: Cells from the human monocytic leukemia cell line THP-1 were grown in RPMI-1640 medium (ATCC, USA) supplemented with 10% fetal bovine serum (FBS) at 37 °C in the presence of 5% CO 2 .

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transduction, Software

    Transwell assay shows that SB reduces THP-1 migration. ( a ) The total number of cells in each well in the lower chamber of the 5 μm-pore transwell were counted and plotted after 24 h of incubation (n = 6 each). Cell counts per well were significantly lower in the FBS + SB group compared with the FBS group. ( b ) SB dose dependency was confirmed by applying different concentrations of SB to the upper chamber of the transwell (n = 4–6). Higher SB concentrations significantly reduced the number of migrating cells. * p < 0.05; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. FBS, fetal bovine serum; SB, sodium benzoate.

    Journal: Scientific Reports

    Article Title: Sodium benzoate attenuates 2,8-dihydroxyadenine nephropathy by inhibiting monocyte/macrophage TNF-α expression

    doi: 10.1038/s41598-023-30056-6

    Figure Lengend Snippet: Transwell assay shows that SB reduces THP-1 migration. ( a ) The total number of cells in each well in the lower chamber of the 5 μm-pore transwell were counted and plotted after 24 h of incubation (n = 6 each). Cell counts per well were significantly lower in the FBS + SB group compared with the FBS group. ( b ) SB dose dependency was confirmed by applying different concentrations of SB to the upper chamber of the transwell (n = 4–6). Higher SB concentrations significantly reduced the number of migrating cells. * p < 0.05; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. FBS, fetal bovine serum; SB, sodium benzoate.

    Article Snippet: Cells from the human monocytic leukemia cell line THP-1 were grown in RPMI-1640 medium (ATCC, USA) supplemented with 10% fetal bovine serum (FBS) at 37 °C in the presence of 5% CO 2 .

    Techniques: Transwell Assay, Migration, Incubation

    SB reduces NF-κB RelB levels of protein, mRNA, and binding to motif DNA sequence. ( a ) Band intensities of NF-κB RelB relative to beta-actin were plotted. In AdCKD kidney, NF-κB RelB protein expression was increased compared to control kidney, which was significantly decreased in AdCKD + SB kidney. ( b ) NF-κB RelB protein expression was increased in LPS stimulated THP-1 cells compared to control cells, which was significantly decreased in SB treated LPS stimulated THP-1 cells. ( c ) mRNA levels of NF-κB RelB in AdCKD kidney were increased compared to control, which was significantly decreased in AdCKD + SB. ( d ) mRNA levels of NF-κB RelB in LPS stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( e ) mRNA levels of NF-κB RelB in TNF-α stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated TNF-α stimulated cells. ( f ) NF-κB RelB binding to motif DNA sequence was increased in AdCKD kidney compared to control, which was significantly decreased in AdCKD + SB kidney. ( g ) NF-κB RelB binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( h ) NF-κB p65 binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was similar levels compared to SB-treated LPS stimulated cells. ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; SB, sodium benzoate.

    Journal: Scientific Reports

    Article Title: Sodium benzoate attenuates 2,8-dihydroxyadenine nephropathy by inhibiting monocyte/macrophage TNF-α expression

    doi: 10.1038/s41598-023-30056-6

    Figure Lengend Snippet: SB reduces NF-κB RelB levels of protein, mRNA, and binding to motif DNA sequence. ( a ) Band intensities of NF-κB RelB relative to beta-actin were plotted. In AdCKD kidney, NF-κB RelB protein expression was increased compared to control kidney, which was significantly decreased in AdCKD + SB kidney. ( b ) NF-κB RelB protein expression was increased in LPS stimulated THP-1 cells compared to control cells, which was significantly decreased in SB treated LPS stimulated THP-1 cells. ( c ) mRNA levels of NF-κB RelB in AdCKD kidney were increased compared to control, which was significantly decreased in AdCKD + SB. ( d ) mRNA levels of NF-κB RelB in LPS stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( e ) mRNA levels of NF-κB RelB in TNF-α stimulated THP-1 cells were increased compared to control, which was significantly decreased in SB-treated TNF-α stimulated cells. ( f ) NF-κB RelB binding to motif DNA sequence was increased in AdCKD kidney compared to control, which was significantly decreased in AdCKD + SB kidney. ( g ) NF-κB RelB binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was significantly decreased in SB-treated LPS stimulated cells in a dose dependent manner. ( h ) NF-κB p65 binding to motif DNA sequence was increased in LPS stimulated THP-1 cells compared to control, which was similar levels compared to SB-treated LPS stimulated cells. ** p < 0.01; *** p < 0.001; ns, not significant. Each bar represents mean ± SEM. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; SB, sodium benzoate.

    Article Snippet: Cells from the human monocytic leukemia cell line THP-1 were grown in RPMI-1640 medium (ATCC, USA) supplemented with 10% fetal bovine serum (FBS) at 37 °C in the presence of 5% CO 2 .

    Techniques: Binding Assay, Sequencing, Expressing