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DSMZ a549 cells
Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) <t>A549</t> cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts
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Images

1) Product Images from "Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿"

Article Title: Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00440-11

Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts
Figure Legend Snippet: Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

Techniques Used: Infection, Mutagenesis

2) Product Images from "Starch-Chitosan Polyplexes: A Versatile Carrier System for Anti-Infectives and Gene Delivery"

Article Title: Starch-Chitosan Polyplexes: A Versatile Carrier System for Anti-Infectives and Gene Delivery

Journal: Polymers

doi: 10.3390/polym10030252

( A ) Representative confocal images of A549 cells transfected with pAmCyan double loaded pAmCyan by using jetPRIME ® as positive control and only cell culture medium as negative control. Transfection was analyzed with CLSM after 48 h, 72 h, and 96 h. Green fluorescence reveals cells successfully transfected with the polyplexes while their morphology remains consistent with non-transfected cells (red: cell membrane; blue: cell nucleus; scale bar 50 μm). ( B ) The transfection efficiency was further quantified using flow cytometry, which indicated the highest amount of transfection after 48 h for pAmCyan double loaded cCP. ( C ) Representative graphs obtained with flow cytometer.
Figure Legend Snippet: ( A ) Representative confocal images of A549 cells transfected with pAmCyan double loaded pAmCyan by using jetPRIME ® as positive control and only cell culture medium as negative control. Transfection was analyzed with CLSM after 48 h, 72 h, and 96 h. Green fluorescence reveals cells successfully transfected with the polyplexes while their morphology remains consistent with non-transfected cells (red: cell membrane; blue: cell nucleus; scale bar 50 μm). ( B ) The transfection efficiency was further quantified using flow cytometry, which indicated the highest amount of transfection after 48 h for pAmCyan double loaded cCP. ( C ) Representative graphs obtained with flow cytometer.

Techniques Used: Transfection, Positive Control, Cell Culture, Negative Control, Confocal Laser Scanning Microscopy, Fluorescence, Flow Cytometry, Cytometry

3) Product Images from "The E3 Ubiquitin Ligase Activity Associated with the Adenoviral E1B-55K-E4orf6 Complex Does Not Require CRM1-Dependent Export ▿"

Article Title: The E3 Ubiquitin Ligase Activity Associated with the Adenoviral E1B-55K-E4orf6 Complex Does Not Require CRM1-Dependent Export ▿

Journal: Journal of Virology

doi: 10.1128/JVI.02368-10

Intracellular localization of E1B-55K, p53, and Mre11 in wt and mutant virus-infected cells. A549 cells were either left uninfected or infected with wt or E1B-55K/E4orf6 mutant viruses at a multiplicity of 20 FFU/cell and were fixed at 24 h p.i. Cells were triple labeled with anti-E1B rat MAb 7C11 (α-E1B), anti-p53 mouse MAb DO-1 (α-p53), and anti-Mre11 rabbit polyclonal antibody pNB-100-142 (α-Mre11) along with Alexa Fluor 488-, 633-, and 555-conjugated secondary antibodies, respectively. Representative anti-E1B (green) (a, f, k, p, u, z, and ae), anti-p53 (blue) (b, g, l, q, v, aa, and af), and anti-Mre11 (red) (c, h, m, r, w, ab, and ag) staining patterns are shown. Overlays of the green, blue, and red images (merge) are also shown (d, i, n, s, x, ac, and ah). Nuclei were visualized using DAPI (blue) (e, j, o, t, y, ad, and ai). Magnification, ×6,300.
Figure Legend Snippet: Intracellular localization of E1B-55K, p53, and Mre11 in wt and mutant virus-infected cells. A549 cells were either left uninfected or infected with wt or E1B-55K/E4orf6 mutant viruses at a multiplicity of 20 FFU/cell and were fixed at 24 h p.i. Cells were triple labeled with anti-E1B rat MAb 7C11 (α-E1B), anti-p53 mouse MAb DO-1 (α-p53), and anti-Mre11 rabbit polyclonal antibody pNB-100-142 (α-Mre11) along with Alexa Fluor 488-, 633-, and 555-conjugated secondary antibodies, respectively. Representative anti-E1B (green) (a, f, k, p, u, z, and ae), anti-p53 (blue) (b, g, l, q, v, aa, and af), and anti-Mre11 (red) (c, h, m, r, w, ab, and ag) staining patterns are shown. Overlays of the green, blue, and red images (merge) are also shown (d, i, n, s, x, ac, and ah). Nuclei were visualized using DAPI (blue) (e, j, o, t, y, ad, and ai). Magnification, ×6,300.

Techniques Used: Mutagenesis, Infection, Labeling, Staining

Effects of E4orf6 NES expression from a lentiviral construct and concomitant infection of A549 cells with wt or mutant viruses on steady-state protein concentrations of p53 and Mre11. E4orf6 NES-expressing A549 cells were infected with wt or mutant viruses at a multiplicity of 20 FFU per cell. Noninfected (m, mock) and infected cells were harvested at the indicated times p.i., and whole-cell extracts were prepared. Proteins (40-μg samples) from each time point were separated on SDS-10% polyacrylamide gels and were subjected to immunoblotting using anti-Mre11 rabbit polyclonal antibody pNB-100-142 and anti-p53 mouse MAb DO-1.
Figure Legend Snippet: Effects of E4orf6 NES expression from a lentiviral construct and concomitant infection of A549 cells with wt or mutant viruses on steady-state protein concentrations of p53 and Mre11. E4orf6 NES-expressing A549 cells were infected with wt or mutant viruses at a multiplicity of 20 FFU per cell. Noninfected (m, mock) and infected cells were harvested at the indicated times p.i., and whole-cell extracts were prepared. Proteins (40-μg samples) from each time point were separated on SDS-10% polyacrylamide gels and were subjected to immunoblotting using anti-Mre11 rabbit polyclonal antibody pNB-100-142 and anti-p53 mouse MAb DO-1.

Techniques Used: Expressing, Construct, Infection, Mutagenesis

Intracellular localization of E1B-55K, E4orf6, and E2A-72K in wt- and NES mutant-infected cells. A549 cells were infected with wt or E1B-55K/E4orf6 mutant viruses at a multiplicity of 20 FFU per cell and were fixed at 20 h p.i. (A) Cells were double labeled with anti-E4orf6 rabbit polyclonal antibody 1807 (α-E4orf6) and anti-E1B-55K mouse MAb 2A6 (α-E1B) as primary antibodies and with FITC- and Texas Red-conjugated secondary antibodies, respectively. Representative anti-E4orf6 (green) (a, d, g, j, m, and p) and anti-E1B (red) (b, e, h, k, n, and q) staining patterns are shown. Overlays of DAPI staining (blue) with the green and red images (merge) are also shown (c, f, i, l, o, and r). Nuclei were visualized using DAPI. Magnification, ×7,600. (B) Cells were labeled with anti-E2A-72K mouse MAb B6-8 and Texas Red-conjugated secondary antibodies (red) (a, c, e, g, i, and k). Representative anti-E2A staining patterns are shown. Overlays of DAPI staining (blue) with the red images (merge) are also shown (b, d, f, h, j, and l). Nuclei were visualized using DAPI. Magnification, ×7,600.
Figure Legend Snippet: Intracellular localization of E1B-55K, E4orf6, and E2A-72K in wt- and NES mutant-infected cells. A549 cells were infected with wt or E1B-55K/E4orf6 mutant viruses at a multiplicity of 20 FFU per cell and were fixed at 20 h p.i. (A) Cells were double labeled with anti-E4orf6 rabbit polyclonal antibody 1807 (α-E4orf6) and anti-E1B-55K mouse MAb 2A6 (α-E1B) as primary antibodies and with FITC- and Texas Red-conjugated secondary antibodies, respectively. Representative anti-E4orf6 (green) (a, d, g, j, m, and p) and anti-E1B (red) (b, e, h, k, n, and q) staining patterns are shown. Overlays of DAPI staining (blue) with the green and red images (merge) are also shown (c, f, i, l, o, and r). Nuclei were visualized using DAPI. Magnification, ×7,600. (B) Cells were labeled with anti-E2A-72K mouse MAb B6-8 and Texas Red-conjugated secondary antibodies (red) (a, c, e, g, i, and k). Representative anti-E2A staining patterns are shown. Overlays of DAPI staining (blue) with the red images (merge) are also shown (b, d, f, h, j, and l). Nuclei were visualized using DAPI. Magnification, ×7,600.

Techniques Used: Mutagenesis, Infection, Labeling, Staining

Effects of NES amino acid changes on the stability of the E1B-55K and/or E4orf6 protein and coimmunoprecipitation of E4orf6 with E1B-55K. (A) Amino acid substitutions in E1B-55K and/or E4orf6 mutant viruses. The NES-specific residues in E1B-55K and E4orf6 are indicated by inverted triangles. Numbers refer to amino acid residues in the wt E1B-55K and E4orf6 proteins from H5 pg 4100. Amino acid changes in the E1B and E4orf6 proteins of H5 pm 4101, H5 pm 4116, and H5 pm 4119 are shown. (B) Steady-state expression levels of E1B-55K and E4orf6 proteins. A549 cells were infected with wt and mutant viruses at a multiplicity of 20 FFU per cell. Cells were harvested at the indicated times p.i., and total-cell extracts were prepared. Proteins (20-μg samples for E1B-55K; 100-μg samples for E4orf6) from each time point were separated by SDS-12% PAGE and were subjected to immunoblotting using anti-E1B-55K (α-E1B) mouse MAb 2A6 or anti-E4orf6 mouse MAb RSA3. (C) Coimmunoprecipitation of E4orf6 with E1B-55K. Whole-cell extracts from infected A549 cells were prepared at 16 h p.i., and samples containing 800 μg of protein (or 2 mg for H5 pm 4116 and H5 pm 4119) were coimmunoprecipitated (Ip) with MAb 2A6 and separated by SDS-12% PAGE, followed by immunoblotting with anti-E4orf6 rabbit polyclonal antibody 1807 (lanes 8 to 14). For analysis of steady-state levels of E1B-55K, E4orf6, and β-actin, proteins (20-μg samples for E1B-55K and β-actin; 100-μg samples for E4orf6) were separated by SDS-12% PAGE and were subjected to immunoblotting using anti-E1B-55K mouse MAb 2A6, anti-E4orf6 rabbit polyclonal antibody 1807, or anti-β-actin mouse MAb AC-15 (lanes 1 to 7).
Figure Legend Snippet: Effects of NES amino acid changes on the stability of the E1B-55K and/or E4orf6 protein and coimmunoprecipitation of E4orf6 with E1B-55K. (A) Amino acid substitutions in E1B-55K and/or E4orf6 mutant viruses. The NES-specific residues in E1B-55K and E4orf6 are indicated by inverted triangles. Numbers refer to amino acid residues in the wt E1B-55K and E4orf6 proteins from H5 pg 4100. Amino acid changes in the E1B and E4orf6 proteins of H5 pm 4101, H5 pm 4116, and H5 pm 4119 are shown. (B) Steady-state expression levels of E1B-55K and E4orf6 proteins. A549 cells were infected with wt and mutant viruses at a multiplicity of 20 FFU per cell. Cells were harvested at the indicated times p.i., and total-cell extracts were prepared. Proteins (20-μg samples for E1B-55K; 100-μg samples for E4orf6) from each time point were separated by SDS-12% PAGE and were subjected to immunoblotting using anti-E1B-55K (α-E1B) mouse MAb 2A6 or anti-E4orf6 mouse MAb RSA3. (C) Coimmunoprecipitation of E4orf6 with E1B-55K. Whole-cell extracts from infected A549 cells were prepared at 16 h p.i., and samples containing 800 μg of protein (or 2 mg for H5 pm 4116 and H5 pm 4119) were coimmunoprecipitated (Ip) with MAb 2A6 and separated by SDS-12% PAGE, followed by immunoblotting with anti-E4orf6 rabbit polyclonal antibody 1807 (lanes 8 to 14). For analysis of steady-state levels of E1B-55K, E4orf6, and β-actin, proteins (20-μg samples for E1B-55K and β-actin; 100-μg samples for E4orf6) were separated by SDS-12% PAGE and were subjected to immunoblotting using anti-E1B-55K mouse MAb 2A6, anti-E4orf6 rabbit polyclonal antibody 1807, or anti-β-actin mouse MAb AC-15 (lanes 1 to 7).

Techniques Used: Mutagenesis, Expressing, Infection, Polyacrylamide Gel Electrophoresis

Effects of amino acid substitutions in the E1B-55K and E4orf6 NES on virus growth, viral DNA accumulation, and viral late protein synthesis. (A) Virus growth. A549 and HeLa cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell and were harvested 72 h p.i., and virus yield was determined by quantitative E2A-72K immunofluorescence staining on 911 cells. The results are reported with Dunnett′s multiple comparison test applied to the log-transformed virus yield (double asterisks indicate significant differences [ P ,
Figure Legend Snippet: Effects of amino acid substitutions in the E1B-55K and E4orf6 NES on virus growth, viral DNA accumulation, and viral late protein synthesis. (A) Virus growth. A549 and HeLa cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell and were harvested 72 h p.i., and virus yield was determined by quantitative E2A-72K immunofluorescence staining on 911 cells. The results are reported with Dunnett′s multiple comparison test applied to the log-transformed virus yield (double asterisks indicate significant differences [ P ,

Techniques Used: Infection, Mutagenesis, Immunofluorescence, Staining, Transformation Assay

Effect of E4orf6 NES expression from a lentiviral construct and concomitant infection of A549 cells with wt and mutant viruses on E1B-55K and/or E4orf6 protein stability, viral late protein synthesis, virus growth, viral late mRNA expression, and the nuclear export efficiency of viral late mRNAs. (A) Steady-state expression levels of E1B-55K and E4orf6. E4orf6 NES-expressing cells were infected with wt and mutant viruses at a multiplicity of 20 FFU per cell. Noninfected (m, mock) and infected cells were harvested at the indicated times p.i., and total-cell extracts were prepared. Proteins (25-μg samples) from each time point were separated by SDS-10% PAGE and were subjected to immunoblotting using anti-E1B-55K mouse MAb 2A6 or anti-E4orf6 rabbit polyclonal antibody 1807. (B) Viral late protein synthesis. Total-cell extracts were prepared at the indicated times postinfection. Proteins (10-μg samples) were separated by SDS-10% PAGE, transferred to nitrocellulose membranes, and probed with anti-Ad5 rabbit polyclonal serum L133 or anti-L4-100K rat MAb 6B10. The bands corresponding to the viral late protein hexon (II) are indicated on the right. (C) Virus growth. E4orf6 NES-expressing cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell and were harvested 72 h p.i., and virus yield was determined by quantitative E2A-72K immunofluorescence staining on 293 cells. The results represent averages for three independent experiments. Error bars indicate the standard deviations of the means. (D) Total amounts of viral late mRNAs L5 and L4. E4orf6 NES-expressing cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell. Steady-state concentrations of both RNAs were determined by real-time PCR at 48 h postinfection. Total L5 and L4 values were corrected using 18S rRNA as an internal control and are expressed relative to the wild-type value. The results represent averages for three independent experiments, each performed in duplicate. Error bars indicate the standard deviations of the means. (E) Nuclear export efficiency of viral late mRNAs L5 and L4. E4orf6 NES-expressing A549 cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell. Steady-state concentrations of both RNAs in the cytoplasm and nucleus were determined by real-time PCR at 48 h postinfection. Raw numbers were corrected using 18S rRNA as an internal control, and the normalized numbers were used to calculate the ratios of cytoplasmic to nuclear L5/L4 mRNAs. The results represent averages for three independent experiments, each performed in duplicate. Error bars indicate the standard deviations of the means.
Figure Legend Snippet: Effect of E4orf6 NES expression from a lentiviral construct and concomitant infection of A549 cells with wt and mutant viruses on E1B-55K and/or E4orf6 protein stability, viral late protein synthesis, virus growth, viral late mRNA expression, and the nuclear export efficiency of viral late mRNAs. (A) Steady-state expression levels of E1B-55K and E4orf6. E4orf6 NES-expressing cells were infected with wt and mutant viruses at a multiplicity of 20 FFU per cell. Noninfected (m, mock) and infected cells were harvested at the indicated times p.i., and total-cell extracts were prepared. Proteins (25-μg samples) from each time point were separated by SDS-10% PAGE and were subjected to immunoblotting using anti-E1B-55K mouse MAb 2A6 or anti-E4orf6 rabbit polyclonal antibody 1807. (B) Viral late protein synthesis. Total-cell extracts were prepared at the indicated times postinfection. Proteins (10-μg samples) were separated by SDS-10% PAGE, transferred to nitrocellulose membranes, and probed with anti-Ad5 rabbit polyclonal serum L133 or anti-L4-100K rat MAb 6B10. The bands corresponding to the viral late protein hexon (II) are indicated on the right. (C) Virus growth. E4orf6 NES-expressing cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell and were harvested 72 h p.i., and virus yield was determined by quantitative E2A-72K immunofluorescence staining on 293 cells. The results represent averages for three independent experiments. Error bars indicate the standard deviations of the means. (D) Total amounts of viral late mRNAs L5 and L4. E4orf6 NES-expressing cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell. Steady-state concentrations of both RNAs were determined by real-time PCR at 48 h postinfection. Total L5 and L4 values were corrected using 18S rRNA as an internal control and are expressed relative to the wild-type value. The results represent averages for three independent experiments, each performed in duplicate. Error bars indicate the standard deviations of the means. (E) Nuclear export efficiency of viral late mRNAs L5 and L4. E4orf6 NES-expressing A549 cells were infected with wt or mutant viruses at a multiplicity of 10 FFU per cell. Steady-state concentrations of both RNAs in the cytoplasm and nucleus were determined by real-time PCR at 48 h postinfection. Raw numbers were corrected using 18S rRNA as an internal control, and the normalized numbers were used to calculate the ratios of cytoplasmic to nuclear L5/L4 mRNAs. The results represent averages for three independent experiments, each performed in duplicate. Error bars indicate the standard deviations of the means.

Techniques Used: Expressing, Construct, Infection, Mutagenesis, Polyacrylamide Gel Electrophoresis, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Effects of leptomycin B treatment on steady-state concentrations and intracellular localization of p53 and Mre11. (A) Steady-state protein levels of p53 and Mre11 in wt-infected and leptomycin B (LMB)-treated cells. A549 cells were either mock infected or infected with the wt virus at a multiplicity of 20 FFU per cell, treated with 20 nM LMB from 12 to 24 (lane 6), 12 to 36 (lane 7), or 24 to 48 (lanes 5 and 8) h p.i., and harvested at the indicated times p.i., and total-cell extracts were prepared. LMB-treated cell lysates (lanes 5 to 8) were compared to untreated controls harvested at the same time points after infection (lanes 1 to 4). Proteins (30-μg samples for p53 and Mre11; 20-μg samples for E1B-55K and E4orf6; 10-μg samples for hexon, fiber, and L4-100K) from each time point were separated by SDS-10% PAGE and were subjected to immunoblotting using anti-p53 mouse MAb DO-1, anti-Mre11 rabbit polyclonal antibody pNB-100-142, anti-E1B-55K mouse MAb 2A6, anti-E4orf6 rabbit polyclonal antibody 1807, anti-Ad5 rabbit polyclonal serum L133, or anti-L4-100K rat MAb 6B10. (B) Intracellular localization of E1B-55K, p53, and Mre11 in wt-infected and LMB-treated cells. A549 cells were either left uninfected or infected with the wt virus at a multiplicity of 20 FFU per cell; either left untreated or treated with 20 nM LMB from 12 to 24 h p.i.; and fixed at 24 h p.i. Cells were triple labeled with anti-E1B rat MAb 7C11 (α-E1B), anti-p53 mouse MAb DO-1 (α-p53), and anti-Mre11 rabbit polyclonal antibody pNB-100-142 (α-Mre11), along with Alexa Fluor 488-, 633-, and 555-conjugated secondary antibodies, respectively. Representative anti-E1B (green) (a, f, k, and p), anti-p53 (blue) (b, g, l, and q), and anti-Mre11 (red) (c, h, m, and r) staining patterns are shown. Overlays of the green, blue, and red images (merge) are also shown (d, i, n, and s). Nuclei were visualized using DAPI (blue) (e, j, o, and t). Magnification, ×6,300.
Figure Legend Snippet: Effects of leptomycin B treatment on steady-state concentrations and intracellular localization of p53 and Mre11. (A) Steady-state protein levels of p53 and Mre11 in wt-infected and leptomycin B (LMB)-treated cells. A549 cells were either mock infected or infected with the wt virus at a multiplicity of 20 FFU per cell, treated with 20 nM LMB from 12 to 24 (lane 6), 12 to 36 (lane 7), or 24 to 48 (lanes 5 and 8) h p.i., and harvested at the indicated times p.i., and total-cell extracts were prepared. LMB-treated cell lysates (lanes 5 to 8) were compared to untreated controls harvested at the same time points after infection (lanes 1 to 4). Proteins (30-μg samples for p53 and Mre11; 20-μg samples for E1B-55K and E4orf6; 10-μg samples for hexon, fiber, and L4-100K) from each time point were separated by SDS-10% PAGE and were subjected to immunoblotting using anti-p53 mouse MAb DO-1, anti-Mre11 rabbit polyclonal antibody pNB-100-142, anti-E1B-55K mouse MAb 2A6, anti-E4orf6 rabbit polyclonal antibody 1807, anti-Ad5 rabbit polyclonal serum L133, or anti-L4-100K rat MAb 6B10. (B) Intracellular localization of E1B-55K, p53, and Mre11 in wt-infected and LMB-treated cells. A549 cells were either left uninfected or infected with the wt virus at a multiplicity of 20 FFU per cell; either left untreated or treated with 20 nM LMB from 12 to 24 h p.i.; and fixed at 24 h p.i. Cells were triple labeled with anti-E1B rat MAb 7C11 (α-E1B), anti-p53 mouse MAb DO-1 (α-p53), and anti-Mre11 rabbit polyclonal antibody pNB-100-142 (α-Mre11), along with Alexa Fluor 488-, 633-, and 555-conjugated secondary antibodies, respectively. Representative anti-E1B (green) (a, f, k, and p), anti-p53 (blue) (b, g, l, and q), and anti-Mre11 (red) (c, h, m, and r) staining patterns are shown. Overlays of the green, blue, and red images (merge) are also shown (d, i, n, and s). Nuclei were visualized using DAPI (blue) (e, j, o, and t). Magnification, ×6,300.

Techniques Used: Infection, Polyacrylamide Gel Electrophoresis, Labeling, Staining

4) Product Images from "Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy"

Article Title: Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy

Journal: Beilstein Journal of Nanotechnology

doi: 10.3762/bjnano.8.130

Confocal microscopic images of A549 cells exposed to 10 µg/mL FD25_Star635 (A) or FD25_Atto647N (B) for 24 h. Images represent the x – y , x – z and y – z sections derived from confocal z -stacks. The orthogonal projections indicate the intracellular location of the particles. The actin cytoskeleton is depicted in cyan, the cis-Golgi network in yellow and silica nanoparticles in magenta.
Figure Legend Snippet: Confocal microscopic images of A549 cells exposed to 10 µg/mL FD25_Star635 (A) or FD25_Atto647N (B) for 24 h. Images represent the x – y , x – z and y – z sections derived from confocal z -stacks. The orthogonal projections indicate the intracellular location of the particles. The actin cytoskeleton is depicted in cyan, the cis-Golgi network in yellow and silica nanoparticles in magenta.

Techniques Used: Derivative Assay

5) Product Images from "KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification"

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification

Journal: Journal of Virology

doi: 10.1128/JVI.01836-15

HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the
Figure Legend Snippet: HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the

Techniques Used: Concentration Assay, Infection

6) Product Images from "KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification"

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification

Journal: Journal of Virology

doi: 10.1128/JVI.01836-15

HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the
Figure Legend Snippet: HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the

Techniques Used: Concentration Assay, Infection

7) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

8) Product Images from "Wittig Derivatization of Sesquiterpenoid Polygodial Leads to Cytostatic Agents with Activity Against Drug Resistant Cancer Cells and Capable of Pyrrolylation of Primary Amines"

Article Title: Wittig Derivatization of Sesquiterpenoid Polygodial Leads to Cytostatic Agents with Activity Against Drug Resistant Cancer Cells and Capable of Pyrrolylation of Primary Amines

Journal: European journal of medicinal chemistry

doi: 10.1016/j.ejmech.2015.08.047

(A) The absense of resistant populations in all 5 cultures tested with analogue 5 and contrasting effects on viability of all cells between 5 and standard chemotherapeutic agents paclitaxel and podophyllotoxin in (B) A549 NSCLC and (C) U87 glioblastoma
Figure Legend Snippet: (A) The absense of resistant populations in all 5 cultures tested with analogue 5 and contrasting effects on viability of all cells between 5 and standard chemotherapeutic agents paclitaxel and podophyllotoxin in (B) A549 NSCLC and (C) U87 glioblastoma

Techniques Used:

9) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

10) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

11) Product Images from "Proteasome-Dependent Degradation of Daxx by the Viral E1B-55K Protein in Human Adenovirus-Infected Cells ▿"

Article Title: Proteasome-Dependent Degradation of Daxx by the Viral E1B-55K Protein in Human Adenovirus-Infected Cells ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00074-10

Degradation of p53, Mre11 and Daxx in different human transformed cell lines. A549, U2OS, and SAOS-2 cells were infected with wild-type (wt) H5 pg 4100 at a multiplicity (moi) of 50 FFU per cell. Cells were harvested at the indicated times postinfection,
Figure Legend Snippet: Degradation of p53, Mre11 and Daxx in different human transformed cell lines. A549, U2OS, and SAOS-2 cells were infected with wild-type (wt) H5 pg 4100 at a multiplicity (moi) of 50 FFU per cell. Cells were harvested at the indicated times postinfection,

Techniques Used: Transformation Assay, Infection

12) Product Images from "Inhibition of Hepatitis E Virus Spread by the Natural Compound Silvestrol"

Article Title: Inhibition of Hepatitis E Virus Spread by the Natural Compound Silvestrol

Journal: Viruses

doi: 10.3390/v10060301

Effect of silvestrol on the intracellular amount of HEV-specific RNA in A549 cells persistently infected with HEV strain 47842c. qPCR analysis using ORF2-specific primers of total RNA isolated from A549 cells persistently infected with HEV; data were normalized to intracellular RPL27 RNA levels. Cells were treated with 2 nM or 50 nM silvestrol, DMSO treatment served as control. n = 3; * p
Figure Legend Snippet: Effect of silvestrol on the intracellular amount of HEV-specific RNA in A549 cells persistently infected with HEV strain 47842c. qPCR analysis using ORF2-specific primers of total RNA isolated from A549 cells persistently infected with HEV; data were normalized to intracellular RPL27 RNA levels. Cells were treated with 2 nM or 50 nM silvestrol, DMSO treatment served as control. n = 3; * p

Techniques Used: Infection, Real-time Polymerase Chain Reaction, Isolation

Repressive effect of silvestrol on release of infectious viral particles from A549 cells persistently infected with hepatitis E virus (HEV) strain 47842c. ( A ) qPCR quantification of extracellular viral RNA in the supernatant derived from silvestrol treated cells (2 nM and 50 nM). The values were referred to the DMSO control; ( B ) Absolute tissue culture infective dose (TCID 50 ) values determined after A549/D3 infection (logarithmic scale); ( C ) Relative TCID 50 values compared to DMSO on the basis of ( B ). n = 3 for all experiments; * p
Figure Legend Snippet: Repressive effect of silvestrol on release of infectious viral particles from A549 cells persistently infected with hepatitis E virus (HEV) strain 47842c. ( A ) qPCR quantification of extracellular viral RNA in the supernatant derived from silvestrol treated cells (2 nM and 50 nM). The values were referred to the DMSO control; ( B ) Absolute tissue culture infective dose (TCID 50 ) values determined after A549/D3 infection (logarithmic scale); ( C ) Relative TCID 50 values compared to DMSO on the basis of ( B ). n = 3 for all experiments; * p

Techniques Used: Infection, Real-time Polymerase Chain Reaction, Derivative Assay

Effect of silvestrol on the genome replication of HEV genotype 3 strain p6Kernow Gaussia luciferase reporter constructs in A549 cells. Intracellular luciferase activity measured in RLU/s normalized to total protein amount after silvestrol treatment (2 and 50 nM) compared to the DMSO-treated cells. n = 3 for all experiments; * p
Figure Legend Snippet: Effect of silvestrol on the genome replication of HEV genotype 3 strain p6Kernow Gaussia luciferase reporter constructs in A549 cells. Intracellular luciferase activity measured in RLU/s normalized to total protein amount after silvestrol treatment (2 and 50 nM) compared to the DMSO-treated cells. n = 3 for all experiments; * p

Techniques Used: Luciferase, Construct, Activity Assay

MVP is upregulated in HEV-infected A549 cells and is downregulated and translocated to the perinuclear region upon 10 nM silvestrol treatment. ( A ) Western Blot analysis of cellular lysates derived from persistently HEV-infected A549 cells as compared to A549/D3. An MVP-specific antibody was used for detection, detection of beta-actin served as loading control. ( B ) Shows the corresponding quantification of ( A ) normalized to beta-actin. n = 4; ** p
Figure Legend Snippet: MVP is upregulated in HEV-infected A549 cells and is downregulated and translocated to the perinuclear region upon 10 nM silvestrol treatment. ( A ) Western Blot analysis of cellular lysates derived from persistently HEV-infected A549 cells as compared to A549/D3. An MVP-specific antibody was used for detection, detection of beta-actin served as loading control. ( B ) Shows the corresponding quantification of ( A ) normalized to beta-actin. n = 4; ** p

Techniques Used: Infection, Western Blot, Derivative Assay

Lactate dehydrogenase (LDH) and PrestoBlue assays to determine silvestrol-mediated cytotoxicity and cytostaticity. ( A , B ) Processed data for LDH-activity in cell-culture supernatants derived from cells treated with 2 or 50 nM silvestrol as compared to respective DMSO controls in ( A ) A549 cells persistently infected with HEV strain 48932c or ( B ) in non-infected A549/D3. ( C , D ) Processed data for redox-metabolic activity in cells treated with 2 or 50 nM silvestrol when compared to respective DMSO controls in ( C ) A549 cells persistently infected with HEV strain 47832c or ( D ) in non-infected A549/D3 cells. * p
Figure Legend Snippet: Lactate dehydrogenase (LDH) and PrestoBlue assays to determine silvestrol-mediated cytotoxicity and cytostaticity. ( A , B ) Processed data for LDH-activity in cell-culture supernatants derived from cells treated with 2 or 50 nM silvestrol as compared to respective DMSO controls in ( A ) A549 cells persistently infected with HEV strain 48932c or ( B ) in non-infected A549/D3. ( C , D ) Processed data for redox-metabolic activity in cells treated with 2 or 50 nM silvestrol when compared to respective DMSO controls in ( C ) A549 cells persistently infected with HEV strain 47832c or ( D ) in non-infected A549/D3 cells. * p

Techniques Used: Activity Assay, Cell Culture, Derivative Assay, Infection

Effects of silvestrol on intracellular HEV capsid protein levels in A549 cells persistently infected with HEV strain 47832c. ( A ) Representative Western Blot of cellular lysates derived from cells treated with either 2 nM or 50 nM silvestrol. DMSO treatment served as control. For detection, a capsid protein specific antiserum and a beta-actin-specific antibody (loading control) were used; ( B ) Relative quantification of Western Blot band intensities normalized to intracellular beta-actin levels referred to the DMSO control. n = 3; * p
Figure Legend Snippet: Effects of silvestrol on intracellular HEV capsid protein levels in A549 cells persistently infected with HEV strain 47832c. ( A ) Representative Western Blot of cellular lysates derived from cells treated with either 2 nM or 50 nM silvestrol. DMSO treatment served as control. For detection, a capsid protein specific antiserum and a beta-actin-specific antibody (loading control) were used; ( B ) Relative quantification of Western Blot band intensities normalized to intracellular beta-actin levels referred to the DMSO control. n = 3; * p

Techniques Used: Infection, Western Blot, Derivative Assay

13) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

14) Product Images from "Comparison of fluorescence-based methods to determine nanoparticle uptake by phagocytes and non-phagocytic cells in vitro"

Article Title: Comparison of fluorescence-based methods to determine nanoparticle uptake by phagocytes and non-phagocytic cells in vitro

Journal: Toxicology

doi: 10.1016/j.tox.2017.01.001

Ratios of DMBM-2/A549 uptake determined by different techniques (a: plate reader, b: by flow cytometry and c: image analysis). Significant differences between the exposure conditions are marked by asterisk.
Figure Legend Snippet: Ratios of DMBM-2/A549 uptake determined by different techniques (a: plate reader, b: by flow cytometry and c: image analysis). Significant differences between the exposure conditions are marked by asterisk.

Techniques Used: Flow Cytometry, Cytometry

Uptake of particles suspended in DMEM + 0% FBS, DMEM + 2% FBS and DMEM + 10% FBS by A549 cells (a) and DMBM-2 macrophages (b) and analyzed by plate reader. Significant differences between DMEM + 0% FBS and media with different FBS content are indicated by asterisks and differences between DMEM + 0% FBS, DMEM + 2% FBS and DMEM + 10% FBS by triangle. Differences in the uptake of one particle compared to all other particles in a given medium are marked with arrow. Data from PPS20 particles were excluded from the analysis due to untypically high variation of the signals (n = 9).
Figure Legend Snippet: Uptake of particles suspended in DMEM + 0% FBS, DMEM + 2% FBS and DMEM + 10% FBS by A549 cells (a) and DMBM-2 macrophages (b) and analyzed by plate reader. Significant differences between DMEM + 0% FBS and media with different FBS content are indicated by asterisks and differences between DMEM + 0% FBS, DMEM + 2% FBS and DMEM + 10% FBS by triangle. Differences in the uptake of one particle compared to all other particles in a given medium are marked with arrow. Data from PPS20 particles were excluded from the analysis due to untypically high variation of the signals (n = 9).

Techniques Used:

Uptake of particles suspended in DMEM + 0% FBS and DMEM + 10% FBS by A549 cells (a) and DMBM-2 macrophages (b) without inhibition and in the presence of 50 mM sodium azide at 4 °C. Evaluation by plate reader with correction of the lower number of cells recovered after the exposure in the inhibitor experiments and normalized to the amount of the particles added to the cells as 100% (n = 3). Inhibition was significant with the exception of PPS20 particles in the exposures to A549 cells.
Figure Legend Snippet: Uptake of particles suspended in DMEM + 0% FBS and DMEM + 10% FBS by A549 cells (a) and DMBM-2 macrophages (b) without inhibition and in the presence of 50 mM sodium azide at 4 °C. Evaluation by plate reader with correction of the lower number of cells recovered after the exposure in the inhibitor experiments and normalized to the amount of the particles added to the cells as 100% (n = 3). Inhibition was significant with the exception of PPS20 particles in the exposures to A549 cells.

Techniques Used: Inhibition

Median of fluorescence in the analysis by flow cytometry in A549 cells (a) and DMBM-2 macrophages (b). Significant differences between the uptakes in DMEM + 0% and DMEM + 10% FBS are marked with asterisks (n = 9).
Figure Legend Snippet: Median of fluorescence in the analysis by flow cytometry in A549 cells (a) and DMBM-2 macrophages (b). Significant differences between the uptakes in DMEM + 0% and DMEM + 10% FBS are marked with asterisks (n = 9).

Techniques Used: Fluorescence, Flow Cytometry, Cytometry

Particle uptake in A549 (a) and DMBM-2 (b) cells seeded at low density (4 × 10 4 cells/chamber) analyzed with image analysis (ImageJ). AFU, arbitrary fluorescence units (n = 4).
Figure Legend Snippet: Particle uptake in A549 (a) and DMBM-2 (b) cells seeded at low density (4 × 10 4 cells/chamber) analyzed with image analysis (ImageJ). AFU, arbitrary fluorescence units (n = 4).

Techniques Used: Fluorescence

Cellular localization of particles after exposure in DMEM + 0% FBS for 24 h. Cells are identified by staining of actin with Alexa Fluor ® 488-phalloidin (green) and particles are seen in red. Nuclei are counterstained with Hoechst 33342 (blue). Co-localization is indicated by yellow color. The intracellular localization of particles is easier to discern in A549 cells because the cells are larger and intensity of the phalloidin staining higher. Scale bar: 50 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Figure Legend Snippet: Cellular localization of particles after exposure in DMEM + 0% FBS for 24 h. Cells are identified by staining of actin with Alexa Fluor ® 488-phalloidin (green) and particles are seen in red. Nuclei are counterstained with Hoechst 33342 (blue). Co-localization is indicated by yellow color. The intracellular localization of particles is easier to discern in A549 cells because the cells are larger and intensity of the phalloidin staining higher. Scale bar: 50 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Techniques Used: Staining

15) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

16) Product Images from "Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿"

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿

Journal:

doi: 10.1128/JVI.02493-08

Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation
Figure Legend Snippet: Binding of wt and mutant L4-100K to eIF4G and viral late mRNAs. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. At 48 h after infection, total cell lysates were prepared and subjected to immunoprecipitation

Techniques Used: Binding Assay, Mutagenesis, Infection, Immunoprecipitation

Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection
Figure Legend Snippet: Protein steady-state levels and L4-100K methylation in A549 cells infected with H5 pg 4100 and H5 pm 4151 viruses. (A) Cells were infected in the absence (AdOx−) or presence (AdOx+) of AdOx and harvested at the indicated times after infection

Techniques Used: Methylation, Infection

Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.
Figure Legend Snippet: Effects of amino acid substitutions in L4-100K on virus growth, viral DNA replication, and late protein synthesis. (A) Virus yield. A549 cells and primary human hepatocytes (PHH) were infected with wt H5 pg 4100 or H5 pm 4151 at the indicated multiplicities.

Techniques Used: Infection

Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection
Figure Legend Snippet: Subcellular localization of wt and mutant L4-100K during productive infection. A549 cells were infected with H5 pg 4100 and H5 pm 4151 viruses at a multiplicity of 10 FFU per cell. (A) Infected cells were fixed at 18 h (a and b) and 48 h (c and d) after infection

Techniques Used: Mutagenesis, Infection

Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested
Figure Legend Snippet: Arginine methylation in RGG boxes modulates hexon binding to L4-100K. (A) A549 cells were infected with H5 pg 4100 and H5 pm 4151 at a multiplicity of 10 FFU per cell in the absence or presence of AdOx (AdOx− and AdOx+, respectively) and harvested

Techniques Used: Methylation, Binding Assay, Infection

17) Product Images from "Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿"

Article Title: Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00440-11

Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts
Figure Legend Snippet: Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

Techniques Used: Infection, Mutagenesis

18) Product Images from "Proteasome-Dependent Degradation of Daxx by the Viral E1B-55K Protein in Human Adenovirus-Infected Cells ▿"

Article Title: Proteasome-Dependent Degradation of Daxx by the Viral E1B-55K Protein in Human Adenovirus-Infected Cells ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00074-10

Degradation of p53, Mre11 and Daxx in different human transformed cell lines. A549, U2OS, and SAOS-2 cells were infected with wild-type (wt) H5 pg 4100 at a multiplicity (moi) of 50 FFU per cell. Cells were harvested at the indicated times postinfection,
Figure Legend Snippet: Degradation of p53, Mre11 and Daxx in different human transformed cell lines. A549, U2OS, and SAOS-2 cells were infected with wild-type (wt) H5 pg 4100 at a multiplicity (moi) of 50 FFU per cell. Cells were harvested at the indicated times postinfection,

Techniques Used: Transformation Assay, Infection

19) Product Images from "Exploiting the 2-Amino-1,3,4-thiadiazole Scaffold To Inhibit Trypanosoma brucei Pteridine Reductase in Support of Early-Stage Drug Discovery "

Article Title: Exploiting the 2-Amino-1,3,4-thiadiazole Scaffold To Inhibit Trypanosoma brucei Pteridine Reductase in Support of Early-Stage Drug Discovery

Journal: ACS Omega

doi: 10.1021/acsomega.7b00473

Summary heat map of the in vitro assays to determine possible liability issues for all synthesized compounds. Assays were performed to assess inhibition of h ERG and five CYP isoforms (1A2, 2C9, 2C19, 2D6, and 3A4), growth inhibition of A549 and W1-38 cell lines, and mitochondrial toxicity at 10 μM. The h DHFR inhibition was determined at 100 μM. The data, expressed as percentage values, were organized by adopting a color code for rapid and intuitive visualization. The ideal compound should have all tiles blue, indicating a safe toxicological profile for progressing into the development pipeline. The cells are colored from blue (100% of cell growth at the tested concentration) to red (0% of cell growth at the tested concentration) for A549 and W1-38 and from blue (0% of inhibitory activity at the tested concentration) to red (100% of inhibitory activity at the tested concentration) for CYP isoforms, h ERG, h DHFR, and mitochondrial toxicity.
Figure Legend Snippet: Summary heat map of the in vitro assays to determine possible liability issues for all synthesized compounds. Assays were performed to assess inhibition of h ERG and five CYP isoforms (1A2, 2C9, 2C19, 2D6, and 3A4), growth inhibition of A549 and W1-38 cell lines, and mitochondrial toxicity at 10 μM. The h DHFR inhibition was determined at 100 μM. The data, expressed as percentage values, were organized by adopting a color code for rapid and intuitive visualization. The ideal compound should have all tiles blue, indicating a safe toxicological profile for progressing into the development pipeline. The cells are colored from blue (100% of cell growth at the tested concentration) to red (0% of cell growth at the tested concentration) for A549 and W1-38 and from blue (0% of inhibitory activity at the tested concentration) to red (100% of inhibitory activity at the tested concentration) for CYP isoforms, h ERG, h DHFR, and mitochondrial toxicity.

Techniques Used: In Vitro, Synthesized, Inhibition, Concentration Assay, Activity Assay

20) Product Images from "KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification"

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification

Journal: Journal of Virology

doi: 10.1128/JVI.01836-15

HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the
Figure Legend Snippet: HAdV induces KAP1 phosphorylation in a virus concentration-dependent manner. (A and B) H1299 cells (A) and A549 cells (B) were infected with wt virus (H5 pg 4100) at a multiplicity of infection of 50 focus-forming units per cell and harvested after the

Techniques Used: Concentration Assay, Infection

Related Articles

Transduction:

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification
Article Snippet: .. To generate KAP1-knockdown cell lines, H1299 and A549 cells were transduced with lentiviral vectors expressing short hairpin RNA (shRNA) targeted to the coding strand sequence 5′-GGAGTTGGATCTCTCAGAA-3′ located at nucleotides 626 to 643 in kap1 . ..

shRNA:

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification
Article Snippet: .. To generate KAP1-knockdown cell lines, H1299 and A549 cells were transduced with lentiviral vectors expressing short hairpin RNA (shRNA) targeted to the coding strand sequence 5′-GGAGTTGGATCTCTCAGAA-3′ located at nucleotides 626 to 643 in kap1 . ..

Infection:

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification
Article Snippet: .. Consistent with these results, KAP1 was also phosphorylated in infected A549 cells, another independent cell line that we tested ( , lanes 6 to 9). .. Moreover, infections with different titers of an HAdV suspension revealed the virus concentration-dependent induction of KAP1 phosphorylation ( ).

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿
Article Snippet: .. A549 cells (3 × 106 ) were collected 48 h after infection, washed with cold PBS, and resuspended in two pellet volumes of polysome lysis buffer containing 100 mM KCl, 5 mM MgCl2 , 10 mM HEPES (pH 7.0), and 0.5% Nonidet P-40 supplemented with 1 mM DTT, 100 units/ml RNase OUT, 0.2 mM phenylmethylsulfonyl fluoride, 1 mg/ml pepstatin A, 5 mg/ml bestatin, and 20 mg/ml leupeptin ( ). .. Resuspended cells were incubated on ice for 5 min and then centrifuged at 16,000 × g for 10 min at 4°C.

Modification:

Article Title: Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿
Article Snippet: .. Primary BRK cells and BRK-derived cell lines (BRK1, AB120, and ABS1) , HEK293 cells , A549 cells (DSMZ ACC107; Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), and the p53-negative human cell line H1299 ( , ) were grown in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 100 U of penicillin, and 100 μg of streptomycin per ml in a 5% CO2 atmosphere at 37°C. .. Mutant Ad5 E1B-55K plasmids were generated from wild-type pE1B-55K , expressing the viral polypeptide under the control of the cytomegalovirus major immediate-early promoter from vector pcDNA3 (Invitrogen) by site-directed mutagenesis with complementary oligonucleotide pairs ( ).

Article Title: The E3 Ubiquitin Ligase Activity Associated with the Adenoviral E1B-55K-E4orf6 Complex Does Not Require CRM1-Dependent Export ▿
Article Snippet: .. HeLa cells (DSMZ ACC 57; Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), A549 cells (DSMZ ACC 107), 911 cells , 293 cells , and W162 ( ) and HEK-293T cells ( ) were grown as monolayer cultures in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS), 100 U of penicillin, and 100 μg of streptomycin per ml under a 5% CO2 atmosphere at 37°C. .. To generate polyclonal E4orf6 NES cells, A549 cells were first transduced with lentiviral particles containing the cDNA of the E4orf6 NES and were afterwards selected and maintained in a medium additionally containing neomycin (250 μg/ml).

Cell Culture:

Article Title: A combined tissue‐engineered/in silico signature tool patient stratification in lung cancer
Article Snippet: .. 5.1 Cell culture HCC827 and A549 cell lines were purchased from DSMZ (Braunschweig, Germany), H441 from ATCC (LGC Standards GmbH – Germany Office, Wesel, Germany). .. A549 and H441 cells were cultured in RPMI + 10% FBS, HCC827 cells in RPMI + 20% FBS.

Expressing:

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification
Article Snippet: .. To generate KAP1-knockdown cell lines, H1299 and A549 cells were transduced with lentiviral vectors expressing short hairpin RNA (shRNA) targeted to the coding strand sequence 5′-GGAGTTGGATCTCTCAGAA-3′ located at nucleotides 626 to 643 in kap1 . ..

Sequencing:

Article Title: KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification
Article Snippet: .. To generate KAP1-knockdown cell lines, H1299 and A549 cells were transduced with lentiviral vectors expressing short hairpin RNA (shRNA) targeted to the coding strand sequence 5′-GGAGTTGGATCTCTCAGAA-3′ located at nucleotides 626 to 643 in kap1 . ..

Lysis:

Article Title: Arginine Methylation of Human Adenovirus Type 5 L4 100-Kilodalton Protein Is Required for Efficient Virus Production ▿
Article Snippet: .. A549 cells (3 × 106 ) were collected 48 h after infection, washed with cold PBS, and resuspended in two pellet volumes of polysome lysis buffer containing 100 mM KCl, 5 mM MgCl2 , 10 mM HEPES (pH 7.0), and 0.5% Nonidet P-40 supplemented with 1 mM DTT, 100 units/ml RNase OUT, 0.2 mM phenylmethylsulfonyl fluoride, 1 mg/ml pepstatin A, 5 mg/ml bestatin, and 20 mg/ml leupeptin ( ). .. Resuspended cells were incubated on ice for 5 min and then centrifuged at 16,000 × g for 10 min at 4°C.

Microscopy:

Article Title: Bright fluorescent silica-nanoparticle probes for high-resolution STED and confocal microscopy
Article Snippet: .. Exposure of cells to NPs and preparation of samples for microscopy Nanoparticle uptake experiments were performed using A549 cells (DSMZ-number: ACC 107) as model for alveolar epithelial type II cells. .. Cells were cultivated in Dulbecco’s modified Eagle medium (DMEM, Gibco, Life Technologies, USA) supplemented with 10% fetal bovine serum (FBS, PAN biotech, Germany) in a humidified incubator (37 °C, 9% CO2 ).

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    DSMZ human alveolar carcinoma cell line a549
    Flow cytometry analysis of H 2 O 2 and NO related to PI staining. <t>A549</t> cells were seeded at 3×10 4 cells/cm 2 and cultured for 24 h. Then, cells were exposed to trypacidin 50 µM or DMSO 1% for 2 h and 24 h. H 2 O 2 and NO production were measured using res pectively H2-DCF-DA, and DAF-FM. IP was used to stain dead cells. Dot-blots are representative of three independent experiments.
    Human Alveolar Carcinoma Cell Line A549, supplied by DSMZ, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human alveolar carcinoma cell line a549/product/DSMZ
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    human alveolar carcinoma cell line a549 - by Bioz Stars, 2020-09
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    93
    DSMZ human lung carcinoma cell line a549
    Influence of ivy preparation and ivy/thyme combination on the β 2 AR binding of <t>A549</t> cells, compared to α-hederin pretreated cells and non-treated control cells. (n ≥ 24 from at least 4 independent experiments, ∗p
    Human Lung Carcinoma Cell Line A549, supplied by DSMZ, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    DSMZ a549 cells
    Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) <t>A549</t> cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts
    A549 Cells, supplied by DSMZ, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Flow cytometry analysis of H 2 O 2 and NO related to PI staining. A549 cells were seeded at 3×10 4 cells/cm 2 and cultured for 24 h. Then, cells were exposed to trypacidin 50 µM or DMSO 1% for 2 h and 24 h. H 2 O 2 and NO production were measured using res pectively H2-DCF-DA, and DAF-FM. IP was used to stain dead cells. Dot-blots are representative of three independent experiments.

    Journal: PLoS ONE

    Article Title: Trypacidin, a Spore-Borne Toxin from Aspergillus fumigatus, Is Cytotoxic to Lung Cells

    doi: 10.1371/journal.pone.0029906

    Figure Lengend Snippet: Flow cytometry analysis of H 2 O 2 and NO related to PI staining. A549 cells were seeded at 3×10 4 cells/cm 2 and cultured for 24 h. Then, cells were exposed to trypacidin 50 µM or DMSO 1% for 2 h and 24 h. H 2 O 2 and NO production were measured using res pectively H2-DCF-DA, and DAF-FM. IP was used to stain dead cells. Dot-blots are representative of three independent experiments.

    Article Snippet: Cell culture conditions The human alveolar carcinoma cell line A549 was obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures (Braunshweig, Germany).

    Techniques: Flow Cytometry, Cytometry, Staining, Cell Culture

    Assessment of ΔΨm. A549 cells were exposed 2 h or 24 h to 10 µM (green line) and 50 µM (blue line) trypacidin or with solely 1% DMSO as negative control (red line). After treatment, cells were stained with DiOC 6 for 20 min. at 37°C. Cells incubated with 40 µM FCCP were used as positive control (brown line). The figure is representative of 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Trypacidin, a Spore-Borne Toxin from Aspergillus fumigatus, Is Cytotoxic to Lung Cells

    doi: 10.1371/journal.pone.0029906

    Figure Lengend Snippet: Assessment of ΔΨm. A549 cells were exposed 2 h or 24 h to 10 µM (green line) and 50 µM (blue line) trypacidin or with solely 1% DMSO as negative control (red line). After treatment, cells were stained with DiOC 6 for 20 min. at 37°C. Cells incubated with 40 µM FCCP were used as positive control (brown line). The figure is representative of 3 independent experiments.

    Article Snippet: Cell culture conditions The human alveolar carcinoma cell line A549 was obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures (Braunshweig, Germany).

    Techniques: Negative Control, Staining, Incubation, Positive Control

    Flow cytometry analysis of the cell cycle. A549 and HBEpC were exposed to 1, 10, 30 and 50 µM trypacidin for 24 h in culture conditions. In the negative control (0 µM trypacidin), cells were exposed to 1% DMSO. Gliotoxin 5 µM was used as positive control to induce sub-G1 particles. The cells were then harvested and analysed for cell cycle as described in the Materials and Methods. The figure shows mean values for 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Trypacidin, a Spore-Borne Toxin from Aspergillus fumigatus, Is Cytotoxic to Lung Cells

    doi: 10.1371/journal.pone.0029906

    Figure Lengend Snippet: Flow cytometry analysis of the cell cycle. A549 and HBEpC were exposed to 1, 10, 30 and 50 µM trypacidin for 24 h in culture conditions. In the negative control (0 µM trypacidin), cells were exposed to 1% DMSO. Gliotoxin 5 µM was used as positive control to induce sub-G1 particles. The cells were then harvested and analysed for cell cycle as described in the Materials and Methods. The figure shows mean values for 3 independent experiments.

    Article Snippet: Cell culture conditions The human alveolar carcinoma cell line A549 was obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures (Braunshweig, Germany).

    Techniques: Flow Cytometry, Cytometry, Negative Control, Positive Control

    Toxicity effect of the trypacidin pathway metabolites on A549 cells. Cell viability and cell lysis were measured using respectively MTT and LDH assay. The cells were exposed for 24 h in culture conditions to each metabolite before measuring cell viability and cell lysis as described in the Materials and Methods. The graph shows the mean values of three independent experiments, excepted for trypacidin for which five experiments were carried out. Negative and positive control corresponds respectively to 1% DMSO and gliotoxin (10 µM for MTT assay and 50 µM for LDH assay). Trypacidin was tested at 50 µM and the other metabolites at 50 and 100 µM. Significant difference from the negative control was analysed with the t-test ( * for p

    Journal: PLoS ONE

    Article Title: Trypacidin, a Spore-Borne Toxin from Aspergillus fumigatus, Is Cytotoxic to Lung Cells

    doi: 10.1371/journal.pone.0029906

    Figure Lengend Snippet: Toxicity effect of the trypacidin pathway metabolites on A549 cells. Cell viability and cell lysis were measured using respectively MTT and LDH assay. The cells were exposed for 24 h in culture conditions to each metabolite before measuring cell viability and cell lysis as described in the Materials and Methods. The graph shows the mean values of three independent experiments, excepted for trypacidin for which five experiments were carried out. Negative and positive control corresponds respectively to 1% DMSO and gliotoxin (10 µM for MTT assay and 50 µM for LDH assay). Trypacidin was tested at 50 µM and the other metabolites at 50 and 100 µM. Significant difference from the negative control was analysed with the t-test ( * for p

    Article Snippet: Cell culture conditions The human alveolar carcinoma cell line A549 was obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures (Braunshweig, Germany).

    Techniques: Lysis, MTT Assay, Lactate Dehydrogenase Assay, Positive Control, Negative Control

    Flow cytometry analysis of ROS and RNS production. A549 cells were seeded at 3×10 4 cells/cm 2 and cultured for 24 h. Then, cells were exposed to trypacidin (green area) 10 µM and 50 µM or DMSO 1% (red line) for 1 h, 2 h and 24 h. H 2 O 2 production was assessed using H2-DCF-DA, O 2•− using DHE, NO using DAF-FM. Menadione was used as control for H2-DCF-DA and DHE, SNAP for DAF-FM (blue line). Peaks are representative of three independent experiments.

    Journal: PLoS ONE

    Article Title: Trypacidin, a Spore-Borne Toxin from Aspergillus fumigatus, Is Cytotoxic to Lung Cells

    doi: 10.1371/journal.pone.0029906

    Figure Lengend Snippet: Flow cytometry analysis of ROS and RNS production. A549 cells were seeded at 3×10 4 cells/cm 2 and cultured for 24 h. Then, cells were exposed to trypacidin (green area) 10 µM and 50 µM or DMSO 1% (red line) for 1 h, 2 h and 24 h. H 2 O 2 production was assessed using H2-DCF-DA, O 2•− using DHE, NO using DAF-FM. Menadione was used as control for H2-DCF-DA and DHE, SNAP for DAF-FM (blue line). Peaks are representative of three independent experiments.

    Article Snippet: Cell culture conditions The human alveolar carcinoma cell line A549 was obtained from the DSMZ-German Collection of Microorganisms and Cell Cultures (Braunshweig, Germany).

    Techniques: Flow Cytometry, Cytometry, Cell Culture

    Influence of ivy preparation and ivy/thyme combination on the β 2 AR binding of A549 cells, compared to α-hederin pretreated cells and non-treated control cells. (n ≥ 24 from at least 4 independent experiments, ∗p

    Journal: Heliyon

    Article Title: A comparative study on the influence of an ivy preparation and an ivy/thyme combination on the β2-adrenergic signal transduction

    doi: 10.1016/j.heliyon.2020.e03960

    Figure Lengend Snippet: Influence of ivy preparation and ivy/thyme combination on the β 2 AR binding of A549 cells, compared to α-hederin pretreated cells and non-treated control cells. (n ≥ 24 from at least 4 independent experiments, ∗p

    Article Snippet: 5.7 Cell Culture The human lung carcinoma cell line A549 obtained from DSMZ was cultivated in RPMI 1640 medium supplemented with 2 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, and 10 % fetal calf serum (all Gibco by Life Technologies, Darmstadt, Germany).

    Techniques: Binding Assay

    Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

    Journal: Journal of Virology

    Article Title: Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿

    doi: 10.1128/JVI.00440-11

    Figure Lengend Snippet: Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

    Article Snippet: Primary BRK cells and BRK-derived cell lines (BRK1, AB120, and ABS1) , HEK293 cells , A549 cells (DSMZ ACC107; Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), and the p53-negative human cell line H1299 ( , ) were grown in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 100 U of penicillin, and 100 μg of streptomycin per ml in a 5% CO2 atmosphere at 37°C.

    Techniques: Infection, Mutagenesis

    ( A ) Representative confocal images of A549 cells transfected with pAmCyan double loaded pAmCyan by using jetPRIME ® as positive control and only cell culture medium as negative control. Transfection was analyzed with CLSM after 48 h, 72 h, and 96 h. Green fluorescence reveals cells successfully transfected with the polyplexes while their morphology remains consistent with non-transfected cells (red: cell membrane; blue: cell nucleus; scale bar 50 μm). ( B ) The transfection efficiency was further quantified using flow cytometry, which indicated the highest amount of transfection after 48 h for pAmCyan double loaded cCP. ( C ) Representative graphs obtained with flow cytometer.

    Journal: Polymers

    Article Title: Starch-Chitosan Polyplexes: A Versatile Carrier System for Anti-Infectives and Gene Delivery

    doi: 10.3390/polym10030252

    Figure Lengend Snippet: ( A ) Representative confocal images of A549 cells transfected with pAmCyan double loaded pAmCyan by using jetPRIME ® as positive control and only cell culture medium as negative control. Transfection was analyzed with CLSM after 48 h, 72 h, and 96 h. Green fluorescence reveals cells successfully transfected with the polyplexes while their morphology remains consistent with non-transfected cells (red: cell membrane; blue: cell nucleus; scale bar 50 μm). ( B ) The transfection efficiency was further quantified using flow cytometry, which indicated the highest amount of transfection after 48 h for pAmCyan double loaded cCP. ( C ) Representative graphs obtained with flow cytometer.

    Article Snippet: A549 cells (human lung carcinoma cell line, No. ACC 107) were obtained from DSMZ GmbH (Braunschweig, Germany).

    Techniques: Transfection, Positive Control, Cell Culture, Negative Control, Confocal Laser Scanning Microscopy, Fluorescence, Flow Cytometry, Cytometry