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primary human small airway epithelial cells hsaecs  (ATCC)


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    Structured Review

    ATCC primary human small airway epithelial cells hsaecs
    A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway <t>epithelial</t> cells <t>(HSAECs)</t> were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.
    Primary Human Small Airway Epithelial Cells Hsaecs, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1413 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway"

    Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

    Journal: Cell Death Discovery

    doi: 10.1038/s41420-026-03122-x

    A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.
    Figure Legend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

    Techniques Used: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker



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    ATCC primary human small airway epithelial cells hsaecs
    A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway <t>epithelial</t> cells <t>(HSAECs)</t> were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.
    Primary Human Small Airway Epithelial Cells Hsaecs, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Epithelix primary hsaecs
    A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway <t>epithelial</t> cells <t>(HSAECs)</t> were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.
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    ATCC hsaec primary small airway epithelial cells
    DSG-PEG2000 turbidity assessment, mucus penetration potential of DSG-PEG2000, and toxicity analysis of DSG-PEG2000 on human respiratory <t>epithelial</t> cells. (A) Turbidity measurement of normal saline, DSG-PEG2000, and chitosan oligosaccharide lactate solutions added to 0.5% porcine mucin II solution, with significance indicated. Error bars represent the standard error of the mean values (* indicates significant difference (p < 0.01), n=3). (B) Visual schematic of artificial mucus layer penetration study (figure created with PowerPoint). (C) Mean absorbance of Nile red in the agarose layer due to mucopenetration. The red line indicates significance between the timepoint 2 hr group. Blue line indicates significance between the timepoint 4 hr group. The green line indicates significance between the timepoint 6 hr group. Error bars represent standard deviation (* indicates significant difference (p < 0.01)). (D) Viability of <t>hSAEC</t> cells following incubation with blank nanoemulsion at concentrations of 20, 40, 60, 80, and 100% (v/v).
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    ATCC hsaec lung epithelial cells
    DSG-PEG2000 turbidity assessment, mucus penetration potential of DSG-PEG2000, and toxicity analysis of DSG-PEG2000 on human respiratory <t>epithelial</t> cells. (A) Turbidity measurement of normal saline, DSG-PEG2000, and chitosan oligosaccharide lactate solutions added to 0.5% porcine mucin II solution, with significance indicated. Error bars represent the standard error of the mean values (* indicates significant difference (p < 0.01), n=3). (B) Visual schematic of artificial mucus layer penetration study (figure created with PowerPoint). (C) Mean absorbance of Nile red in the agarose layer due to mucopenetration. The red line indicates significance between the timepoint 2 hr group. Blue line indicates significance between the timepoint 4 hr group. The green line indicates significance between the timepoint 6 hr group. Error bars represent standard deviation (* indicates significant difference (p < 0.01)). (D) Viability of <t>hSAEC</t> cells following incubation with blank nanoemulsion at concentrations of 20, 40, 60, 80, and 100% (v/v).
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    hsaecs  (ATCC)
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    ATCC hsaecs
    DSG-PEG2000 turbidity assessment, mucus penetration potential of DSG-PEG2000, and toxicity analysis of DSG-PEG2000 on human respiratory <t>epithelial</t> cells. (A) Turbidity measurement of normal saline, DSG-PEG2000, and chitosan oligosaccharide lactate solutions added to 0.5% porcine mucin II solution, with significance indicated. Error bars represent the standard error of the mean values (* indicates significant difference (p < 0.01), n=3). (B) Visual schematic of artificial mucus layer penetration study (figure created with PowerPoint). (C) Mean absorbance of Nile red in the agarose layer due to mucopenetration. The red line indicates significance between the timepoint 2 hr group. Blue line indicates significance between the timepoint 4 hr group. The green line indicates significance between the timepoint 6 hr group. Error bars represent standard deviation (* indicates significant difference (p < 0.01)). (D) Viability of <t>hSAEC</t> cells following incubation with blank nanoemulsion at concentrations of 20, 40, 60, 80, and 100% (v/v).
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    ATCC primary hsaec
    Secreted DARS2 from airway epithelia binds to macrophages. (a) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (b) quantitation of DARS2 in the supernatant from primary human small <t>airway</t> <t>epithelial</t> cells <t>(HSAEC)</t> treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (c) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (d) quantitation of DARS2 in the supernatant from human alveolar macrophages (HAM) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (e) Schematic of adoptive media transfer from BEAS‐2Bs to recipient PMA‐stimulated THP1‐macrophages. BEAS2Bs were transfected with FLAG‐tagged empty vector (EV) or DARS2‐Flag plasmid and media was transferred to THP‐1 cells. (f) Shown are levels of ectopically expressed DARS2 in THP1 cell lysates (each lane is a biologically independent replicate, repeated for n = 2). (g) Immunofluorescence of THP1‐macrophages receiving media from donor BEAS‐2B cells transfected with DARS2‐FLAG as in (e, f) showing co‐localization of DARS2‐Flag with the actin filament marker, phalloidin (arrows, merge panels) Scale bar = 20 μm (repeated for n = 2). (h) IL18 , (i) TNF , (j) CXCL1 mRNA levels by RT‐qPCR analysis in THP1‐macrophages 24 h after receiving media from donor BEAS‐2B cells transfected with EV‐FLAG or DARS2‐FLAG ( n = 8 biological replicates/group).
    Primary Hsaec, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC primary small airway epithelial cells hsaecs
    Secreted DARS2 from airway epithelia binds to macrophages. (a) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (b) quantitation of DARS2 in the supernatant from primary human small <t>airway</t> <t>epithelial</t> cells <t>(HSAEC)</t> treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (c) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (d) quantitation of DARS2 in the supernatant from human alveolar macrophages (HAM) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (e) Schematic of adoptive media transfer from BEAS‐2Bs to recipient PMA‐stimulated THP1‐macrophages. BEAS2Bs were transfected with FLAG‐tagged empty vector (EV) or DARS2‐Flag plasmid and media was transferred to THP‐1 cells. (f) Shown are levels of ectopically expressed DARS2 in THP1 cell lysates (each lane is a biologically independent replicate, repeated for n = 2). (g) Immunofluorescence of THP1‐macrophages receiving media from donor BEAS‐2B cells transfected with DARS2‐FLAG as in (e, f) showing co‐localization of DARS2‐Flag with the actin filament marker, phalloidin (arrows, merge panels) Scale bar = 20 μm (repeated for n = 2). (h) IL18 , (i) TNF , (j) CXCL1 mRNA levels by RT‐qPCR analysis in THP1‐macrophages 24 h after receiving media from donor BEAS‐2B cells transfected with EV‐FLAG or DARS2‐FLAG ( n = 8 biological replicates/group).
    Primary Small Airway Epithelial Cells Hsaecs, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

    Journal: Cell Death Discovery

    Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

    doi: 10.1038/s41420-026-03122-x

    Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

    Article Snippet: Primary human small airway epithelial cells (HSAECs) (PCS-301-010) were obtained from ATCC and cultured in Airway Epithelial Cell Basal Medium (ATCC) containing a Bronchial Epithelial Cell Growth Kit (ATCC) and penicillin-streptomycin (100 U/mL, 100 μg/mL).

    Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker

    DSG-PEG2000 turbidity assessment, mucus penetration potential of DSG-PEG2000, and toxicity analysis of DSG-PEG2000 on human respiratory epithelial cells. (A) Turbidity measurement of normal saline, DSG-PEG2000, and chitosan oligosaccharide lactate solutions added to 0.5% porcine mucin II solution, with significance indicated. Error bars represent the standard error of the mean values (* indicates significant difference (p < 0.01), n=3). (B) Visual schematic of artificial mucus layer penetration study (figure created with PowerPoint). (C) Mean absorbance of Nile red in the agarose layer due to mucopenetration. The red line indicates significance between the timepoint 2 hr group. Blue line indicates significance between the timepoint 4 hr group. The green line indicates significance between the timepoint 6 hr group. Error bars represent standard deviation (* indicates significant difference (p < 0.01)). (D) Viability of hSAEC cells following incubation with blank nanoemulsion at concentrations of 20, 40, 60, 80, and 100% (v/v).

    Journal: Frontiers in Immunology

    Article Title: Development and evaluation of an inhalable nanoemulsion system for enhancing NK cell function against osteosarcoma pulmonary metastases

    doi: 10.3389/fimmu.2026.1772375

    Figure Lengend Snippet: DSG-PEG2000 turbidity assessment, mucus penetration potential of DSG-PEG2000, and toxicity analysis of DSG-PEG2000 on human respiratory epithelial cells. (A) Turbidity measurement of normal saline, DSG-PEG2000, and chitosan oligosaccharide lactate solutions added to 0.5% porcine mucin II solution, with significance indicated. Error bars represent the standard error of the mean values (* indicates significant difference (p < 0.01), n=3). (B) Visual schematic of artificial mucus layer penetration study (figure created with PowerPoint). (C) Mean absorbance of Nile red in the agarose layer due to mucopenetration. The red line indicates significance between the timepoint 2 hr group. Blue line indicates significance between the timepoint 4 hr group. The green line indicates significance between the timepoint 6 hr group. Error bars represent standard deviation (* indicates significant difference (p < 0.01)). (D) Viability of hSAEC cells following incubation with blank nanoemulsion at concentrations of 20, 40, 60, 80, and 100% (v/v).

    Article Snippet: Recombinant human (rh)IL-2 and rhIL-15 were obtained from the Biological Resources Branch (National Cancer Institute, Frederick, MD) and recombinant murine IL-15 was purchased from R&D Systems (Minneapolis, MN). hSAEC primary small airway epithelial cells (Cat #PCS-301-010), NK-92 human NK cell lymphoma (Cat #CRL-2407), MG63 human osteosarcoma (Cat #CRL-1427), and K7M2 murine osteosarcoma (Cat #CRL-2836) cell lines were all obtained from ATCC (Manassas, VA).

    Techniques: Saline, Standard Deviation, Incubation

    Secreted DARS2 from airway epithelia binds to macrophages. (a) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (b) quantitation of DARS2 in the supernatant from primary human small airway epithelial cells (HSAEC) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (c) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (d) quantitation of DARS2 in the supernatant from human alveolar macrophages (HAM) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (e) Schematic of adoptive media transfer from BEAS‐2Bs to recipient PMA‐stimulated THP1‐macrophages. BEAS2Bs were transfected with FLAG‐tagged empty vector (EV) or DARS2‐Flag plasmid and media was transferred to THP‐1 cells. (f) Shown are levels of ectopically expressed DARS2 in THP1 cell lysates (each lane is a biologically independent replicate, repeated for n = 2). (g) Immunofluorescence of THP1‐macrophages receiving media from donor BEAS‐2B cells transfected with DARS2‐FLAG as in (e, f) showing co‐localization of DARS2‐Flag with the actin filament marker, phalloidin (arrows, merge panels) Scale bar = 20 μm (repeated for n = 2). (h) IL18 , (i) TNF , (j) CXCL1 mRNA levels by RT‐qPCR analysis in THP1‐macrophages 24 h after receiving media from donor BEAS‐2B cells transfected with EV‐FLAG or DARS2‐FLAG ( n = 8 biological replicates/group).

    Journal: Physiological Reports

    Article Title: Secreted mitochondrial aspartyl‐ tRNA synthetase ( DARS2 ) regulates TNFα signaling

    doi: 10.14814/phy2.70627

    Figure Lengend Snippet: Secreted DARS2 from airway epithelia binds to macrophages. (a) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (b) quantitation of DARS2 in the supernatant from primary human small airway epithelial cells (HSAEC) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (c) Immunoreactive levels of DARS2 in the cell lysate (top) and supernatant (bottom) and (d) quantitation of DARS2 in the supernatant from human alveolar macrophages (HAM) treated with Pam, IL‐1β or increasing concentrations of TNFα (repeated for n = 3 representative blots shown). (e) Schematic of adoptive media transfer from BEAS‐2Bs to recipient PMA‐stimulated THP1‐macrophages. BEAS2Bs were transfected with FLAG‐tagged empty vector (EV) or DARS2‐Flag plasmid and media was transferred to THP‐1 cells. (f) Shown are levels of ectopically expressed DARS2 in THP1 cell lysates (each lane is a biologically independent replicate, repeated for n = 2). (g) Immunofluorescence of THP1‐macrophages receiving media from donor BEAS‐2B cells transfected with DARS2‐FLAG as in (e, f) showing co‐localization of DARS2‐Flag with the actin filament marker, phalloidin (arrows, merge panels) Scale bar = 20 μm (repeated for n = 2). (h) IL18 , (i) TNF , (j) CXCL1 mRNA levels by RT‐qPCR analysis in THP1‐macrophages 24 h after receiving media from donor BEAS‐2B cells transfected with EV‐FLAG or DARS2‐FLAG ( n = 8 biological replicates/group).

    Article Snippet: BEAS‐2B (Human Bronchial Epithelial cells, ATCC, catalog no. CRL‐35588), THP‐1 (Tohoku Hospital Pediatrics‐1, an acute myeloid leukemia line used as a model of monocytes, ATCC, catalog no. TIB‐202), primary HSAEC (human small airway epithelial cells, ATCC, catalog no. PCS‐301‐010), and primary HBEC (human bronchial epithelial cells, ATCC, catalog no. PCS‐300‐010) were used for this study.

    Techniques: Quantitation Assay, Transfection, Plasmid Preparation, Immunofluorescence, Marker, Quantitative RT-PCR