a549 cells line  (Millipore)


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    Name:
    A549 Cell Line human
    Description:

    Catalog Number:
    cb_86012804
    Price:
    None
    Applications:
    Human A549 cell line has been used:. to study the ability of 2-(2,4-dihydroxyphenyl)thieno-1,3-thiazin-4-one (BChTT) to inhibit cancer cell proliferation and identify mechanisms involved in its actions. to study the effects of insulin and insulin-like growth factor 1 (IGF1) on proliferation and apoptosis in A549 cells. to culture A549 cells in different media over a long term and identify conditions for promoting alveolar type II differentiation characteristics in A549 cells
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    Structured Review

    Millipore a549 cells line
    A549 Cell Line human

    https://www.bioz.com/result/a549 cells line/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    a549 cells line - by Bioz Stars, 2020-09
    99/100 stars

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    Images

    1) Product Images from "Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells"

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    Cell cycle distribution of A549 cells line and A549 tumor spheres on effects of chemotherapeutic drugs
    Figure Legend Snippet: Cell cycle distribution of A549 cells line and A549 tumor spheres on effects of chemotherapeutic drugs

    Techniques Used:

    Isolation and identification of LCSCs. A. Three types of colonies formed by A549 cells. (400×). B. Immunostaining of stem cell markers in secondary spheres. Nuclei were stained using DAPI (blue). LCM, scale bar = 25 10, 50, 75, 10, 30 μm
    Figure Legend Snippet: Isolation and identification of LCSCs. A. Three types of colonies formed by A549 cells. (400×). B. Immunostaining of stem cell markers in secondary spheres. Nuclei were stained using DAPI (blue). LCM, scale bar = 25 10, 50, 75, 10, 30 μm

    Techniques Used: Isolation, Immunostaining, Staining, Laser Capture Microdissection

    Enhanced expression of GST-π, MRP1 and LRP in A549 spheres. A. Western-blot of GST-π, MRP1 and LRP in HEB, A549 cells and A549 spheres. B. Immunofluorescence of GST-π expression in HEB, A549 cells and A549 spheres using confocal
    Figure Legend Snippet: Enhanced expression of GST-π, MRP1 and LRP in A549 spheres. A. Western-blot of GST-π, MRP1 and LRP in HEB, A549 cells and A549 spheres. B. Immunofluorescence of GST-π expression in HEB, A549 cells and A549 spheres using confocal

    Techniques Used: Expressing, Western Blot, Immunofluorescence

    Effect of the IC50 concentration of Cisplatin (DDP) and Gemcitabine (GEM) on the cell cycle profile of A549 cells line and A549 spheres. A. A549 cells line; B. A549 cells line/DDP; C. A549 cells line/GEM; D. A549 cancer spheres; E. A549 cancer spheres/DDP;
    Figure Legend Snippet: Effect of the IC50 concentration of Cisplatin (DDP) and Gemcitabine (GEM) on the cell cycle profile of A549 cells line and A549 spheres. A. A549 cells line; B. A549 cells line/DDP; C. A549 cells line/GEM; D. A549 cancer spheres; E. A549 cancer spheres/DDP;

    Techniques Used: Concentration Assay

    Measurement of the sensitivity of A549 cancer spheres and A549 cells line to anticancer drugs (DDP and GEM). A. The lethal doses (IC50) of DDP (left) and GEM (right) in A549 cancer spheres and A549 cells line, The data shown represent the mean ±
    Figure Legend Snippet: Measurement of the sensitivity of A549 cancer spheres and A549 cells line to anticancer drugs (DDP and GEM). A. The lethal doses (IC50) of DDP (left) and GEM (right) in A549 cancer spheres and A549 cells line, The data shown represent the mean ±

    Techniques Used:

    A549 cancer spheres have high express CD133 and more self renewal capability. A. The morphology of tumor spheres. a. Primary tumor spheres formed in the CSM medium. b. Secondary spheres derived from single parental cells of primary tumor spheres under
    Figure Legend Snippet: A549 cancer spheres have high express CD133 and more self renewal capability. A. The morphology of tumor spheres. a. Primary tumor spheres formed in the CSM medium. b. Secondary spheres derived from single parental cells of primary tumor spheres under

    Techniques Used: Derivative Assay

    Related Articles

    MTT Assay:

    Article Title: Combination Therapy With Polydeoxyribonucleotide and Pirfenidone Alleviates Symptoms of Acute Respiratory Distress Syndrome in Human Lung Epithelial A549 Cells
    Article Snippet: .. MTT Assay To determine the viability, A549 cells were plated at a density of 1×104 cells/well in a 96-well plate, as the previously described method [ , ]. .. For the induction of ARDS environment, 1-μg/mL LPS (Escherichia coli serotype 026:B6; Sigma-Aldrich Chemical Co., St. Louis, MO, USA) and 5-ng/mL TGF-β (Kolon Pharm., Seoul, Korea) were treated.

    Cell Culture:

    Article Title: Emodin Attenuates Bleomycin-Induced Pulmonary Fibrosis via Anti-Inflammatory and Anti-Oxidative Activities in Rats
    Article Snippet: .. Cell culture The human A549 cell line (Sigma Aldrich, Shanghai, China) was obtained and maintained in DMEM (Thermo Fisher Scientific) complemented with 10% FBS (Hyclone, Logan, Utah, USA) in 95% air and 5% CO2 at 37°C. ..

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis
    Article Snippet: .. 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C. ..

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes
    Article Snippet: .. Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere. .. Cells were washed before and after incubation with washing buffer [4.5 g/L glucose and 5 mM MgCl2 in Dulbecco’s PBS with calcium chloride and magnesium chloride (Sigma-Aldrich)].

    Article Title: Melatonin Enhances Palladium-Nanoparticle-Induced Cytotoxicity and Apoptosis in Human Lung Epithelial Adenocarcinoma Cells A549 and H1229
    Article Snippet: .. A549 cells were seeded into wells of 24 well plates at a density of 5 × 104 cells per well and cultured for 24 h. After washing twice with phosphate-buffered saline (PBS), fresh medium containing 2.5 µM of PdNPs, 0.75 mM of MLT, 2.5 µM of PdNPs combined with 0.75 mM of MLT, or 5 µM of DOX was added and cells were incubated for 24 h. Cells were supplemented with 20 μM DCFH2-DA, and incubation continued for 30 min at 37 °C. .. Cells were then rinsed with PBS (2 mL of PBS was added to each well) and the fluorescence intensity was determined using a Gemini EM spectrofluorometer (Molecular Devices, Sunnyvale, CA, USA) at an excitation wavelength of 485 nm and an emission wavelength of 530 nm.

    Concentration Assay:

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis
    Article Snippet: .. 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C. ..

    Incubation:

    Article Title: Melatonin Enhances Palladium-Nanoparticle-Induced Cytotoxicity and Apoptosis in Human Lung Epithelial Adenocarcinoma Cells A549 and H1229
    Article Snippet: .. A549 cells were seeded into wells of 24 well plates at a density of 5 × 104 cells per well and cultured for 24 h. After washing twice with phosphate-buffered saline (PBS), fresh medium containing 2.5 µM of PdNPs, 0.75 mM of MLT, 2.5 µM of PdNPs combined with 0.75 mM of MLT, or 5 µM of DOX was added and cells were incubated for 24 h. Cells were supplemented with 20 μM DCFH2-DA, and incubation continued for 30 min at 37 °C. .. Cells were then rinsed with PBS (2 mL of PBS was added to each well) and the fluorescence intensity was determined using a Gemini EM spectrofluorometer (Molecular Devices, Sunnyvale, CA, USA) at an excitation wavelength of 485 nm and an emission wavelength of 530 nm.

    other:

    Article Title: Combination Therapy With Polydeoxyribonucleotide and Pirfenidone Alleviates Symptoms of Acute Respiratory Distress Syndrome in Human Lung Epithelial A549 Cells
    Article Snippet: A549 cells was washed in phosphate buffered saline to removed media.

    Article Title: Glucocorticoids rapidly inhibit cell migration through a novel, non-transcriptional HDAC6 pathway
    Article Snippet: A549 cells were pre-conditioned to 100 nM dex or vehicle control (DMSO) for 48 h (37°C/5% CO2 ).

    Modification:

    Article Title: Small-Molecule Screening Using a Whole-Cell Viral Replication Reporter Gene Assay Identifies 2-{[2-(Benzoylamino)Benzoyl]Amino}-Benzoic Acid as a Novel Antiadenoviral Compound ▿
    Article Snippet: .. A549 cells (oat cell carcinoma from the human lung; alveolar basal epithelial cells) were grown in Dulbecco modified Eagle medium (DMEM; Sigma-Aldrich, St. Louis, MO) containing 0.75 g of NaHCO3 /liter, 20 mM HEPES (EuroClone, Milan, Italy), penicillin G (100 IU/ml), and streptomycin sulfate (100 μg/ml) combined (1× PEST; Gibco, Carlsbad, CA), and 5% fetal bovine serum (FBS; Gibco) at 37°C. ..

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis
    Article Snippet: .. 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C. ..

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes
    Article Snippet: .. Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere. .. Cells were washed before and after incubation with washing buffer [4.5 g/L glucose and 5 mM MgCl2 in Dulbecco’s PBS with calcium chloride and magnesium chloride (Sigma-Aldrich)].

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  • 84
    Millipore a549 cell line human
    Transmission of SARS-CoV-2 genome into human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) through EVs. A , Schematic depiction of study design. Nsp1 indicates non-structural protein 1; Nsp12, non-structural protein 12; E, envelope protein; N, nucleocapsid protein. B , Expression of SARS-Cov-2 genes in <t>A549</t> lung epithelial cells. A549 cells were infected with indicated lentiviral particles for 48 hours and mRNA levels were measured by qRT-PCR (left) (n=3, mean± S.D) and semiquantitative PCR (right). *P
    A549 Cell Line Human, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a549 cell line human/product/Millipore
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    a549 cell line human - by Bioz Stars, 2020-09
    84/100 stars
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    94
    Millipore a549 cells
    Autophagy adapter protein p62 controls the production of active AMPs by interacting with AMP precursors. (A B) Silencing P62 affected the active peptide levels of hBD1 and LL37. <t>A549</t> cells were transiently transfected with si-control or si-SQSTM1 to silence P62 and then infected with BCG. The expression of hBD1 and LL37 was detected using western blotting and real-time PCR. (C) The autophagic level of A549 cells affected the co-localization level of hBD1/LL37 precursor and p62-positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 3-MA and rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1/LL37 precursor, p62, and GFP-LC3, labeled using Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. (D) Direct interaction could be observed between p62 and hCAP18. A549 cells were infected with BCG for 24 and 48 h. The interaction between p62 and hCAP18 was detected by Co-IP with anti-p62 antibody followed by western blotting with anti-p62 and anti-hCAP18 antibodies. Data are expressed as the means ± standard deviation (s.d.) * p
    A549 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 343 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a549 cells/product/Millipore
    Average 94 stars, based on 343 article reviews
    Price from $9.99 to $1999.99
    a549 cells - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    Transmission of SARS-CoV-2 genome into human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) through EVs. A , Schematic depiction of study design. Nsp1 indicates non-structural protein 1; Nsp12, non-structural protein 12; E, envelope protein; N, nucleocapsid protein. B , Expression of SARS-Cov-2 genes in A549 lung epithelial cells. A549 cells were infected with indicated lentiviral particles for 48 hours and mRNA levels were measured by qRT-PCR (left) (n=3, mean± S.D) and semiquantitative PCR (right). *P

    Journal: bioRxiv

    Article Title: Detection of Synthetic Viral RNA Fragments in Human iPSC-Cardiomyocytes following Treatment with Precipitated Extracellular Vesicles from SARS-CoV-2 Coding-Sequence-Overexpressing Lung Epithelial Cells

    doi: 10.1101/2020.05.14.093583

    Figure Lengend Snippet: Transmission of SARS-CoV-2 genome into human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) through EVs. A , Schematic depiction of study design. Nsp1 indicates non-structural protein 1; Nsp12, non-structural protein 12; E, envelope protein; N, nucleocapsid protein. B , Expression of SARS-Cov-2 genes in A549 lung epithelial cells. A549 cells were infected with indicated lentiviral particles for 48 hours and mRNA levels were measured by qRT-PCR (left) (n=3, mean± S.D) and semiquantitative PCR (right). *P

    Article Snippet: Isolation of extracellular vesicles Supernatant of A549 cells was collected at 48 hours after transduction with lentivirus for isolation of EVs.

    Techniques: Transmission Assay, Derivative Assay, Expressing, Infection, Quantitative RT-PCR, Polymerase Chain Reaction

    Effect of curcumin and/or LPS on cell viability and apoptosis of A549 cells. A549 cells were treated with curcumin (10 µM) and/or LPS (1 µg/ml) for different time. (A) Cell viability was monitored by CellTiter 96® AQueous One Solution Cell Proliferation Assay. Data is presented as mean ± Standard error of mean of six data points. (B) Cells were fixed and then stained with propidium iodide (50 µg/ml) and Annexin V-FITC. The DNA contents were analyzed by flow cytometry. Representative result from three independent experiments is presented. x- and y-axis present DNA content and cell number, respectively. (C) Cells (in %) undergoing early- apoptotic or late-apoptotic phase were calculated using Flowing software.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: Effect of curcumin and/or LPS on cell viability and apoptosis of A549 cells. A549 cells were treated with curcumin (10 µM) and/or LPS (1 µg/ml) for different time. (A) Cell viability was monitored by CellTiter 96® AQueous One Solution Cell Proliferation Assay. Data is presented as mean ± Standard error of mean of six data points. (B) Cells were fixed and then stained with propidium iodide (50 µg/ml) and Annexin V-FITC. The DNA contents were analyzed by flow cytometry. Representative result from three independent experiments is presented. x- and y-axis present DNA content and cell number, respectively. (C) Cells (in %) undergoing early- apoptotic or late-apoptotic phase were calculated using Flowing software.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Proliferation Assay, Staining, Flow Cytometry, Software

    Global A549 proteome and common proteins identified in curcumin and/or LPS treated cells. (A) Venn diagram representing the total number of proteins identified in CURI, LPSI and LPSCUR experiments. (B) Subnetwork of 778 common genes (696 were mapped to the database with 65 linker genes) to all 3 experiments (CURI, LPSI and LPSCUR). (C) Venn diagram representing the proteins identified in CURI, LPSI and LPSCUR experiments with similar abundance in expression. (D) Subnetwork of 427 common genes (385 genes were mapped to database with 47 linker genes) with similar abundance in all the 3 experiments are shown. Color coded modules in which the interaction of the genes are highest based on the modularity. The functions of the genes are predicted by Reactome FI plugin in Cytoscape 3.4.0. The circled maps of the subnetworks involve different biological processes. Linker genes are shown by diamond shaped nodes.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: Global A549 proteome and common proteins identified in curcumin and/or LPS treated cells. (A) Venn diagram representing the total number of proteins identified in CURI, LPSI and LPSCUR experiments. (B) Subnetwork of 778 common genes (696 were mapped to the database with 65 linker genes) to all 3 experiments (CURI, LPSI and LPSCUR). (C) Venn diagram representing the proteins identified in CURI, LPSI and LPSCUR experiments with similar abundance in expression. (D) Subnetwork of 427 common genes (385 genes were mapped to database with 47 linker genes) with similar abundance in all the 3 experiments are shown. Color coded modules in which the interaction of the genes are highest based on the modularity. The functions of the genes are predicted by Reactome FI plugin in Cytoscape 3.4.0. The circled maps of the subnetworks involve different biological processes. Linker genes are shown by diamond shaped nodes.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Expressing

    Schematic diagram showing probable mechanism of curcumin action on LPS treated A549 cells. Curcumin (C) inhibits activation of several proteins and the ones marked with dark red oval are identified in this study.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: Schematic diagram showing probable mechanism of curcumin action on LPS treated A549 cells. Curcumin (C) inhibits activation of several proteins and the ones marked with dark red oval are identified in this study.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Activation Assay

    Deregulated global proteome in the A549 cells treated with curcumin and/or LPS. (A) Treatment of curcumin (10 µM) or LPS (1 µg/ml) for 24 hours alters proteome levels in A549 cell lines. Subnetwork in (B) Functional categorization and distribution of deregulated proteins in CURI (305) and LPSI (346) experiments. (C) Mitochondrial proteins showing inverse relationship in both CURI and LPSI experiments. (D) Simultaneous treatment of curcumin (10 µM) and LPS (1 µg/ml) for 48 hours induced proteome change in A549 cell lines. (E) Functional categorization and the distribution of LPSCUR (372) differentially expressed proteins to extract the information of their known functions.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: Deregulated global proteome in the A549 cells treated with curcumin and/or LPS. (A) Treatment of curcumin (10 µM) or LPS (1 µg/ml) for 24 hours alters proteome levels in A549 cell lines. Subnetwork in (B) Functional categorization and distribution of deregulated proteins in CURI (305) and LPSI (346) experiments. (C) Mitochondrial proteins showing inverse relationship in both CURI and LPSI experiments. (D) Simultaneous treatment of curcumin (10 µM) and LPS (1 µg/ml) for 48 hours induced proteome change in A549 cell lines. (E) Functional categorization and the distribution of LPSCUR (372) differentially expressed proteins to extract the information of their known functions.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Functional Assay

    Experimental strategy adopted in this study to understand the anti-inflammatory role of curcumin in A549 cells treated with LPS. Experimental design used to analyze altered proteome in A549 cells treated with curcumin (10 μM) and LPS (1 μg/ml) in SILAC experiments and other function assays used in this study including Mycobacteria infection.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: Experimental strategy adopted in this study to understand the anti-inflammatory role of curcumin in A549 cells treated with LPS. Experimental design used to analyze altered proteome in A549 cells treated with curcumin (10 μM) and LPS (1 μg/ml) in SILAC experiments and other function assays used in this study including Mycobacteria infection.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Infection

    H37Rv-GFP infected A549 cells, treated with curcumin and/or LPS, for 24 hours at multiple of infectivity (1:5) showed successful Mtb internalization and varied bacterial load. (A) Representative confocal microscopy images of cells stained with DAPI (blue, 1 µg/ml) to stain nucleus. Scale: 20 µm and calculation from average of 5 different focal points in triplicates for each experiment. Additional images are available in Figure S9A, S9B and S9C. (B) Colony forming unit (CFU) counts at different conditions. * at 95 % confidence and ** at 99 % confidence. (C) Expression of BID and AIFM1 proteins as probed using Western blot analysis. GAPDH was used as loading control and relative band intensity are presented.

    Journal: bioRxiv

    Article Title: Anti-inflammatory role of curcumin in Lipopolysaccharide treated A549 cells at global proteome level and on mycobacterial infection

    doi: 10.1101/721100

    Figure Lengend Snippet: H37Rv-GFP infected A549 cells, treated with curcumin and/or LPS, for 24 hours at multiple of infectivity (1:5) showed successful Mtb internalization and varied bacterial load. (A) Representative confocal microscopy images of cells stained with DAPI (blue, 1 µg/ml) to stain nucleus. Scale: 20 µm and calculation from average of 5 different focal points in triplicates for each experiment. Additional images are available in Figure S9A, S9B and S9C. (B) Colony forming unit (CFU) counts at different conditions. * at 95 % confidence and ** at 99 % confidence. (C) Expression of BID and AIFM1 proteins as probed using Western blot analysis. GAPDH was used as loading control and relative band intensity are presented.

    Article Snippet: Monitoring Mitochondrial distribution in treated A549 cells using Confocal Microscopy A549 cells were grown overnight on 18 mm coverslip in twelve well plates at 37 °C with 5% CO2 and incubated with curcumin/LPS/both for appropriate durations.

    Techniques: Infection, Confocal Microscopy, Staining, Expressing, Western Blot

    Expression of an EV-associated protein in A549 cell lysates. Dot blot analyses of CD81 in A549 cell lysates following LPS treatment (0.1 µg/mL, 1 µg/mL, and 10 µg/mL).

    Journal: bioRxiv

    Article Title: Lipopolysaccharide administration alters extracellular vesicles in human lung-cancer cells and mice

    doi: 10.1101/2020.04.17.046367

    Figure Lengend Snippet: Expression of an EV-associated protein in A549 cell lysates. Dot blot analyses of CD81 in A549 cell lysates following LPS treatment (0.1 µg/mL, 1 µg/mL, and 10 µg/mL).

    Article Snippet: A549 cells were seeded independently in 96-well tissue culture plates (10,000 cells/well) and maintained in culture for 24 hrs prior to treatment.

    Techniques: Expressing, Dot Blot

    LPS treatment alters EVs from A549 cells. (A) Mean sizes and (B) particle concentrations (per mL) were determined for A549-derived EVs following LPS treatment using Nanosight Tracking Analysis. ELISAs of A549-derived EVs were used to examine expressions of (C) LAMP-1, and (D) RRP44/DIS3 proteins. Mean fold change ± SEM data are from a total of five experiments. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.

    Journal: bioRxiv

    Article Title: Lipopolysaccharide administration alters extracellular vesicles in human lung-cancer cells and mice

    doi: 10.1101/2020.04.17.046367

    Figure Lengend Snippet: LPS treatment alters EVs from A549 cells. (A) Mean sizes and (B) particle concentrations (per mL) were determined for A549-derived EVs following LPS treatment using Nanosight Tracking Analysis. ELISAs of A549-derived EVs were used to examine expressions of (C) LAMP-1, and (D) RRP44/DIS3 proteins. Mean fold change ± SEM data are from a total of five experiments. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.

    Article Snippet: A549 cells were seeded independently in 96-well tissue culture plates (10,000 cells/well) and maintained in culture for 24 hrs prior to treatment.

    Techniques: Derivative Assay

    Autophagy adapter protein p62 controls the production of active AMPs by interacting with AMP precursors. (A B) Silencing P62 affected the active peptide levels of hBD1 and LL37. A549 cells were transiently transfected with si-control or si-SQSTM1 to silence P62 and then infected with BCG. The expression of hBD1 and LL37 was detected using western blotting and real-time PCR. (C) The autophagic level of A549 cells affected the co-localization level of hBD1/LL37 precursor and p62-positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 3-MA and rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1/LL37 precursor, p62, and GFP-LC3, labeled using Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. (D) Direct interaction could be observed between p62 and hCAP18. A549 cells were infected with BCG for 24 and 48 h. The interaction between p62 and hCAP18 was detected by Co-IP with anti-p62 antibody followed by western blotting with anti-p62 and anti-hCAP18 antibodies. Data are expressed as the means ± standard deviation (s.d.) * p

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: Autophagy adapter protein p62 controls the production of active AMPs by interacting with AMP precursors. (A B) Silencing P62 affected the active peptide levels of hBD1 and LL37. A549 cells were transiently transfected with si-control or si-SQSTM1 to silence P62 and then infected with BCG. The expression of hBD1 and LL37 was detected using western blotting and real-time PCR. (C) The autophagic level of A549 cells affected the co-localization level of hBD1/LL37 precursor and p62-positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 3-MA and rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1/LL37 precursor, p62, and GFP-LC3, labeled using Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. (D) Direct interaction could be observed between p62 and hCAP18. A549 cells were infected with BCG for 24 and 48 h. The interaction between p62 and hCAP18 was detected by Co-IP with anti-p62 antibody followed by western blotting with anti-p62 and anti-hCAP18 antibodies. Data are expressed as the means ± standard deviation (s.d.) * p

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Transfection, Infection, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Stable Transfection, Fluorescence, Confocal Microscopy, Labeling, Co-Immunoprecipitation Assay, Standard Deviation

    Antimicrobial peptides hBD1 and LL37 are highly expressed in MTB-infected lung epithelial cells and exhibit anti-MTB activity. (A) High level expression of hBD1 and LL37 was detected in A549 cells. The clls were infected with BCG for 48 h and the seven AMP mRNAs were evaluated by real-time PCR. (B C) hBD1 (B) and LL37 (C) mRNA expression in BCG-infected A549 cells was detected at specified time points by real-time PCR. (D) The active peptides of hBD1 and LL37 were detected at specified time points and evaluated by western blotting. (E F) High level expression of hBD1 and LL37 mRNA in PBMCs of patients with TB. (G) High level expression of LL37 was detected in human alveolar type II pneumocytes from patients with TB via immunohistochemistry. Micrograph shows strong LL37 immunostaining in human alveolar type II pneumocytes from patients with TB compared to that in healthy tissue donors and patients with pneumoconiosis (magnification x200). (H-J) Silencing of hBD1 or/and LL37 decreased intracellular BCG killing in A549 cells. The intracellular viable bacilli were determined by CFU assays at 72 h. Survival rate was calculated compared with that at 0 h. Data are expressed as the means ± standard deviation (s.d.). * p

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: Antimicrobial peptides hBD1 and LL37 are highly expressed in MTB-infected lung epithelial cells and exhibit anti-MTB activity. (A) High level expression of hBD1 and LL37 was detected in A549 cells. The clls were infected with BCG for 48 h and the seven AMP mRNAs were evaluated by real-time PCR. (B C) hBD1 (B) and LL37 (C) mRNA expression in BCG-infected A549 cells was detected at specified time points by real-time PCR. (D) The active peptides of hBD1 and LL37 were detected at specified time points and evaluated by western blotting. (E F) High level expression of hBD1 and LL37 mRNA in PBMCs of patients with TB. (G) High level expression of LL37 was detected in human alveolar type II pneumocytes from patients with TB via immunohistochemistry. Micrograph shows strong LL37 immunostaining in human alveolar type II pneumocytes from patients with TB compared to that in healthy tissue donors and patients with pneumoconiosis (magnification x200). (H-J) Silencing of hBD1 or/and LL37 decreased intracellular BCG killing in A549 cells. The intracellular viable bacilli were determined by CFU assays at 72 h. Survival rate was calculated compared with that at 0 h. Data are expressed as the means ± standard deviation (s.d.). * p

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Infection, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Immunostaining, Standard Deviation

    The autophagic level of A549 cells affected the co-localization level of hBD1 precursor and p62 positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with 3-MA and Rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1, p62 and GFP-LC3, marked with Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The autophagic level of A549 cells affected the co-localization level of hBD1 precursor and p62 positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with 3-MA and Rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1, p62 and GFP-LC3, marked with Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection, Confocal Microscopy

    Autophagy influences the active peptide levels of hBD1 and LL37. (A) The autophagic level of A549 cells affected the active peptides level of hBD1 and LL37. A549 cells were pretreated with 4 μM rapamycin for 6 h and 5 mM 3-MA for 2 h and then infected with BCG for 24 h. The active peptide levels of hBD1 and LL37 were evaluated by western blotting. (B) The autophagic level of A549 cells did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (C D) Starvation-induced autophagy promoted the production of active hBD1 and LL37. A549 cells were cultured with EBSS culture medium at various time points. The expression of hBD1 and LL37 was detected by western blotting and real-time PCR. (E F) Silencing ATG 5 and 7 disturbed the production of active hBD1 and LL37. A549 cells were transfected with 100 nM si-control or siRNA for ATG 5 and 7 for 72 h and infected with BCG. The expression of hBD1 and LL37 was detected by western blotting and real-time PCR. (G) The autophagic level of A549 cells affected the co-localization level of hBD1/LL37 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)– tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 or LL37 precursor (hBD1/LL37 precursor-mCherry) were pretreated with rapamycin and 3-MA and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1/LL37 precursor and autophagosomes was detected by confocal microscopy. Data are expressed as the means ± standard deviation (s.d.) * p

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: Autophagy influences the active peptide levels of hBD1 and LL37. (A) The autophagic level of A549 cells affected the active peptides level of hBD1 and LL37. A549 cells were pretreated with 4 μM rapamycin for 6 h and 5 mM 3-MA for 2 h and then infected with BCG for 24 h. The active peptide levels of hBD1 and LL37 were evaluated by western blotting. (B) The autophagic level of A549 cells did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (C D) Starvation-induced autophagy promoted the production of active hBD1 and LL37. A549 cells were cultured with EBSS culture medium at various time points. The expression of hBD1 and LL37 was detected by western blotting and real-time PCR. (E F) Silencing ATG 5 and 7 disturbed the production of active hBD1 and LL37. A549 cells were transfected with 100 nM si-control or siRNA for ATG 5 and 7 for 72 h and infected with BCG. The expression of hBD1 and LL37 was detected by western blotting and real-time PCR. (G) The autophagic level of A549 cells affected the co-localization level of hBD1/LL37 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)– tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 or LL37 precursor (hBD1/LL37 precursor-mCherry) were pretreated with rapamycin and 3-MA and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1/LL37 precursor and autophagosomes was detected by confocal microscopy. Data are expressed as the means ± standard deviation (s.d.) * p

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Infection, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture, Expressing, Transfection, Stable Transfection, Fluorescence, Confocal Microscopy, Standard Deviation

    The autophagic level of A549 cells affected the co-localization level of hBD1 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with rapamycin and 3-MA as described above and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1 precursor and autophagosomes was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The autophagic level of A549 cells affected the co-localization level of hBD1 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with rapamycin and 3-MA as described above and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of hBD1 precursor and autophagosomes was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection, Confocal Microscopy

    hBD1 and LL37 production was affected by the function of lysosomes. (A) The formation of autolysosomes affected the active peptide levels of hBD1 and LL37. A549 cells were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. The active peptide levels of hBD1 and LL37 were evaluated using western blotting. (B) The formation of autolysosomes did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (C) The function of lysosomes affected the active peptide levels of hBD1 and LL37. A549 cells were pretreated with 10 mg/mL NH4Cl and infected with BCG for 48 h. The active peptide levels of hBD1 and LL37 were evaluated by western blotting. (D) The function of lysosomes did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (E F) The formation of autolysosomes did not affect the co-localization rates between hBD1/LL37 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 or LL37 precursor (hBD1/LL37 precursor-mCherry) were pretreated and infected with BCG as described as (A). Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. Co-localization of hBD1/LL37 precursor and autolysosomes was detected by confocal microscopy. Data are expressed as the means ± standard deviation (s.d.) * p

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: hBD1 and LL37 production was affected by the function of lysosomes. (A) The formation of autolysosomes affected the active peptide levels of hBD1 and LL37. A549 cells were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. The active peptide levels of hBD1 and LL37 were evaluated using western blotting. (B) The formation of autolysosomes did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (C) The function of lysosomes affected the active peptide levels of hBD1 and LL37. A549 cells were pretreated with 10 mg/mL NH4Cl and infected with BCG for 48 h. The active peptide levels of hBD1 and LL37 were evaluated by western blotting. (D) The function of lysosomes did not affect the mRNA level of hBD1 and LL37. A549 cells were pretreated and infected with BCG as described above. The mRNA levels of hBD1 and LL37 were evaluated using real-time PCR. (E F) The formation of autolysosomes did not affect the co-localization rates between hBD1/LL37 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 or LL37 precursor (hBD1/LL37 precursor-mCherry) were pretreated and infected with BCG as described as (A). Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. Co-localization of hBD1/LL37 precursor and autolysosomes was detected by confocal microscopy. Data are expressed as the means ± standard deviation (s.d.) * p

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Infection, Western Blot, Real-time Polymerase Chain Reaction, Stable Transfection, Expressing, Fluorescence, Confocal Microscopy, Standard Deviation

    The autophagic level of A549 cells affected the co-localization level of LL37 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with rapamycin and 3-MA as described above and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of LL37 precursor and autophagosomes was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The autophagic level of A549 cells affected the co-localization level of LL37 precursor and autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with rapamycin and 3-MA as described above and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of LL37 precursor and autophagosomes was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection, Confocal Microscopy

    The formation of autolysosomes could not affect the co-localization rates between LL37 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. The function of lysosomes could not affect the co-localization rates between LL37 precursor and autolysosomes. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The formation of autolysosomes could not affect the co-localization rates between LL37 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. The function of lysosomes could not affect the co-localization rates between LL37 precursor and autolysosomes. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection

    Autophagy-related production of antimicrobial peptides is a novel mechanism of autophagy-mediated BCG killing. (A-C) Silencing of hBD1 (A) or/and LL37 (B C) weakened the autophagic killing of intracellular BCG. A549 cells were transfected with si-NC, Si-DEFB1, or/and si-CAMP for 24 h then infected with BCG (MOI = 5) for 1 h. In the rapamycin (Rapa) group the cells were pretreated by 4 μM rapamycin for 6 h prior to BCG infection. The intracellular viable bacilli were determined by CFU assays at 72 h. Survival rate was calculated compared with that at 0 h. Data are expressed as the means ± standard deviation (s.d.) * p

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: Autophagy-related production of antimicrobial peptides is a novel mechanism of autophagy-mediated BCG killing. (A-C) Silencing of hBD1 (A) or/and LL37 (B C) weakened the autophagic killing of intracellular BCG. A549 cells were transfected with si-NC, Si-DEFB1, or/and si-CAMP for 24 h then infected with BCG (MOI = 5) for 1 h. In the rapamycin (Rapa) group the cells were pretreated by 4 μM rapamycin for 6 h prior to BCG infection. The intracellular viable bacilli were determined by CFU assays at 72 h. Survival rate was calculated compared with that at 0 h. Data are expressed as the means ± standard deviation (s.d.) * p

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Transfection, Infection, Standard Deviation

    The autophagic level of A549 cells affected the co-localization level of LL37 precursor and p62 positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 3-MA and Rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of LL37, p62 and GFP-LC3, marked with Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The autophagic level of A549 cells affected the co-localization level of LL37 precursor and p62 positive autophagosomes. A549 cells stably expressing green fluorescent protein (GFP)-tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged LL37 precursor (LL37 precursor-mCherry) were pretreated with 3-MA and Rapamycin and infected with BCG as described above. GFP-LC3 puncta ( > 1 μm) were observed and counted under confocal microscopy. Co-localization of LL37, p62 and GFP-LC3, marked with Alexa Fluor 647-coupled antibody against p62, was detected by confocal microscopy. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection, Confocal Microscopy

    The formation of autolysosomes could not affect the co-localization rates between hBD1 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. The function of lysosomes could not affect the co-localization rates between hBD1 precursor and autolysosomes. These experiments were performed independently at least thrice with similar results.

    Journal: bioRxiv

    Article Title: Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

    doi: 10.1101/2020.02.21.959361

    Figure Lengend Snippet: The formation of autolysosomes could not affect the co-localization rates between hBD1 precursor and autolysosomes. A549 cells stably expressing GFP–tagged LC3 (GFP-LC3) and mCherry Fluorescence Protein (mCherry)-tagged hBD1 precursor (hBD1 precursor-mCherry) were pretreated with 0.1 mM Baf A1 or/and 4 μM rapamycin and infected with BCG for 48 h. Lysosome was labelled with lyso tracker, GFP-LC3 ( > 1 μm) and lyso tracker double positive puncta were determined as autolysosomes. The function of lysosomes could not affect the co-localization rates between hBD1 precursor and autolysosomes. These experiments were performed independently at least thrice with similar results.

    Article Snippet: A549 cells were infected with GFP-LC3 lentivirus (17-10193; Millipore, Bedford, MA, USA) according to manufacturer instruction.

    Techniques: Stable Transfection, Expressing, Fluorescence, Infection