a549 cells  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH a549 cells
    A549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    a549 cells  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH a549 cells
    A549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    human lung adenocarcinoma cell line a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH human lung adenocarcinoma cell line a549
    Human Lung Adenocarcinoma Cell Line A549, supplied by CLS Cell Lines Service GmbH, 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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    a 549 cells  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH a 549 cells
    A 549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    cell lines a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH cell lines a549
    Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. <t>A549</t> cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.
    Cell Lines A549, supplied by CLS Cell Lines Service GmbH, 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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    1) Product Images from "The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling"

    Article Title: The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling

    Journal: Vaccines

    doi: 10.3390/vaccines11040768

    Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. A549 cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.
    Figure Legend Snippet: Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. A549 cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.

    Techniques Used: Activation Assay, Binding Assay, Western Blot

    Time-course accumulation of pEGFR and pERK1/2 in A549 cells treated with full length Spike 1 protein and its RBD. Cells were treated or not with 2.5 μg/mL full length Spike protein ( A ) or 5 μg/mL RBD ( B ) at different times as indicated (5, 15, 30 and 60 min) and the protein levels of pEGFR and pERK1/2 and their respective total levels were determined by western blot. The treatment with EGF (5 μg/mL) was carried out for 15 min.
    Figure Legend Snippet: Time-course accumulation of pEGFR and pERK1/2 in A549 cells treated with full length Spike 1 protein and its RBD. Cells were treated or not with 2.5 μg/mL full length Spike protein ( A ) or 5 μg/mL RBD ( B ) at different times as indicated (5, 15, 30 and 60 min) and the protein levels of pEGFR and pERK1/2 and their respective total levels were determined by western blot. The treatment with EGF (5 μg/mL) was carried out for 15 min.

    Techniques Used: Western Blot

    Blockade of EGFR inhibits SARS-CoV-2 spike 1-mediated activation of AKT in A549 cells. A549 were first pre-treated with AG1478 (10 μM) for 15 min prior to treatment with EGF ( A , B ), full-length spike 1 protein or spike RBD protein ( B ) as described in . Whole cell extracts were resolved on SDS-PAGE and protein levels were determined by western blot.
    Figure Legend Snippet: Blockade of EGFR inhibits SARS-CoV-2 spike 1-mediated activation of AKT in A549 cells. A549 were first pre-treated with AG1478 (10 μM) for 15 min prior to treatment with EGF ( A , B ), full-length spike 1 protein or spike RBD protein ( B ) as described in . Whole cell extracts were resolved on SDS-PAGE and protein levels were determined by western blot.

    Techniques Used: Activation Assay, SDS Page, Western Blot

    SARS-CoV-2 full length spike 1 protein and its RBD promote survivin expression in A549 cells. A549 cells were treated or not with EGF (5 μg/mL) ( A , B ), full-length spike 1 protein (2.5 μg/mL) ( A ) or spike RBD protein (5 μg/mL) ( B ) at different times (5, 15, 30 and 60 min) as indicated and the protein level of survivin was determined by Western blot. ( C ) A549 cells were first pre-treated or not with AG1478 (10 μM) for 15 min prior to treatment with EGF (5 μg/mL) or spike RBD protein (5 μg/mL) and the protein level of survivin was determined as described above.
    Figure Legend Snippet: SARS-CoV-2 full length spike 1 protein and its RBD promote survivin expression in A549 cells. A549 cells were treated or not with EGF (5 μg/mL) ( A , B ), full-length spike 1 protein (2.5 μg/mL) ( A ) or spike RBD protein (5 μg/mL) ( B ) at different times (5, 15, 30 and 60 min) as indicated and the protein level of survivin was determined by Western blot. ( C ) A549 cells were first pre-treated or not with AG1478 (10 μM) for 15 min prior to treatment with EGF (5 μg/mL) or spike RBD protein (5 μg/mL) and the protein level of survivin was determined as described above.

    Techniques Used: Expressing, Western Blot

    cell lines a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH cell lines a549
    Cell Lines A549, supplied by CLS Cell Lines Service GmbH, 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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    human non small cell lung carcinoma a549 cells  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH human non small cell lung carcinoma a549 cells
    Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from <t>A549</t> and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).
    Human Non Small Cell Lung Carcinoma A549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    1) Product Images from "Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models"

    Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2022.1045521

    Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from A549 and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).
    Figure Legend Snippet: Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from A549 and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).

    Techniques Used: Derivative Assay, In Vitro, Staining, Expressing, Quantitative RT-PCR

    Characterization of cell lines (continued). Relative expression of mesenchymal and epithelial markers cytokeratin (A) and vimentin (B) (proteins of intermediate filaments family) and activated protein kinases phospho-ERK and phospho-AKT (C) as determined by the Western blot method. Representative pictures from more than 3 independent experiments are shown. Coomassie-stained polyacrylamide gels serve as loading controls. Numbers indicate cell lines as in <xref ref-type= Figure 1 ; the letter A stands for the cell line A549. " title="... the letter A stands for the cell line A549. " property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Characterization of cell lines (continued). Relative expression of mesenchymal and epithelial markers cytokeratin (A) and vimentin (B) (proteins of intermediate filaments family) and activated protein kinases phospho-ERK and phospho-AKT (C) as determined by the Western blot method. Representative pictures from more than 3 independent experiments are shown. Coomassie-stained polyacrylamide gels serve as loading controls. Numbers indicate cell lines as in Figure 1 ; the letter A stands for the cell line A549.

    Techniques Used: Expressing, Western Blot, Staining

    Phosphorylation of ERK1/2 and AKT in lung cancer-derived cell lines after cisplatin treatment. (A) The dynamics of AKT and ERK phosphorylation in A549 cells. (B) Changes in AKT and ERK phosphorylation in lung tumor-derived cell lines after 6 hours of cisplatin treatment. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. The concentration of cisplatin was 90 µM.
    Figure Legend Snippet: Phosphorylation of ERK1/2 and AKT in lung cancer-derived cell lines after cisplatin treatment. (A) The dynamics of AKT and ERK phosphorylation in A549 cells. (B) Changes in AKT and ERK phosphorylation in lung tumor-derived cell lines after 6 hours of cisplatin treatment. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. The concentration of cisplatin was 90 µM.

    Techniques Used: Derivative Assay, Western Blot, Staining, Concentration Assay

    Dependence of the interplay between ERK and AKT signaling pathways on extracellular contacts. (A) Focal adhesion kinase inhibitor PF573228 (+Fi; 10 µM) attenuates the MEK kinase inhibitor selumetinib-induced increase of AKT phosphorylation in control and cisplatin-treated lung cancer-derived cells, except A549 cells. (B) PF573228 prevents the AKT inhibitor capivasertib-induced increase in ERK phosphorylation in control and cisplatin-treated lung cancer-derived cells. (C) Cells grown in suspension (Susp) under agitation, in contrast to adherent cells (Adh), do not show alternative kinase activation after the treatment with inhibitors. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), +cis – cisplatin (90 µM), CAP – capivasertib (10 µM), AKTi – AKT inhibitor VIII (10 µM).
    Figure Legend Snippet: Dependence of the interplay between ERK and AKT signaling pathways on extracellular contacts. (A) Focal adhesion kinase inhibitor PF573228 (+Fi; 10 µM) attenuates the MEK kinase inhibitor selumetinib-induced increase of AKT phosphorylation in control and cisplatin-treated lung cancer-derived cells, except A549 cells. (B) PF573228 prevents the AKT inhibitor capivasertib-induced increase in ERK phosphorylation in control and cisplatin-treated lung cancer-derived cells. (C) Cells grown in suspension (Susp) under agitation, in contrast to adherent cells (Adh), do not show alternative kinase activation after the treatment with inhibitors. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), +cis – cisplatin (90 µM), CAP – capivasertib (10 µM), AKTi – AKT inhibitor VIII (10 µM).

    Techniques Used: Derivative Assay, Activation Assay, Western Blot, Staining

    a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH a549
    a , b Immunofluorescence images of <t>A549</t> cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.
    A549, supplied by CLS Cell Lines Service GmbH, 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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    1) Product Images from "HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells"

    Article Title: HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells

    Journal: Nature Communications

    doi: 10.1038/s41467-022-32537-0

    a , b Immunofluorescence images of A549 cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.
    Figure Legend Snippet: a , b Immunofluorescence images of A549 cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.

    Techniques Used: Immunofluorescence, Western Blot

    a Representative immunofluorescence images showing the effect of doxycycline (DOX) induced overexpression (−DOX/+DOX) of HnRNPK (green) on global dsRNA as detected using J2 antibody (red) in A549 cells. Panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar on the images is 50 um. Bar graph to the right shows the number of J2 signals in HnRNPK non-induced (−DOX (−D); n = 81 cells) and HnRNPK induced (DOX+(+D); n = 53 cells) cells from three replicates quantified as RNA spots per cell using Imaris spot detection tools. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-tailed unpaired t -test. b Immunoblot showing the levels of HnRNPK and IER3 in −DOX (−D) and +DOX (+D) A549 cells. GAPDH was used as an internal loading control. The similar results were observed in an independent experiment. Histograms showing the relative band intensities of the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates. c RNAscope images of DOX (−D/+D) induced HnRNPK cells showing IER3 (red) and IER3-AS1 (green) in control (−D) and hnRNPK overexpressed (+D) cells. Panel to the right showing magnified images of the merged panel indicated by white box. Yellow arrows depict colocalized IER3/IER3-AS1 dsRNA (yellow dots). Indicative scale bar on the images is 50 um. d Bar graph showing the number of IER3-AS1 and IER3 RNA signals per cell in control (−D; n = 80 cells) and hnRNPK overexpressed (+D; n = 121 cells) cells quantified using Imaris spot detection tools from three replicates. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test. e Dot plots showing the average number of colocalized signals of IER3-AS1/IER3 (dsRNA) per cell in control (−D) and hnRNPK overexpressed (+D) cells. A total of 150 cells were manually counted from three independent replicates. Data are presented as mean values + / SEM. Statistical significance was calculated by Two-tailed unpaired t -test. For figures ( a , d and e ) * p ≤ 0.05; ** p ≤ 0.005; *** p ≤ 0.0005. f Model explaining the role of HnRNPK in the regulation of global dsRNA.
    Figure Legend Snippet: a Representative immunofluorescence images showing the effect of doxycycline (DOX) induced overexpression (−DOX/+DOX) of HnRNPK (green) on global dsRNA as detected using J2 antibody (red) in A549 cells. Panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar on the images is 50 um. Bar graph to the right shows the number of J2 signals in HnRNPK non-induced (−DOX (−D); n = 81 cells) and HnRNPK induced (DOX+(+D); n = 53 cells) cells from three replicates quantified as RNA spots per cell using Imaris spot detection tools. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-tailed unpaired t -test. b Immunoblot showing the levels of HnRNPK and IER3 in −DOX (−D) and +DOX (+D) A549 cells. GAPDH was used as an internal loading control. The similar results were observed in an independent experiment. Histograms showing the relative band intensities of the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates. c RNAscope images of DOX (−D/+D) induced HnRNPK cells showing IER3 (red) and IER3-AS1 (green) in control (−D) and hnRNPK overexpressed (+D) cells. Panel to the right showing magnified images of the merged panel indicated by white box. Yellow arrows depict colocalized IER3/IER3-AS1 dsRNA (yellow dots). Indicative scale bar on the images is 50 um. d Bar graph showing the number of IER3-AS1 and IER3 RNA signals per cell in control (−D; n = 80 cells) and hnRNPK overexpressed (+D; n = 121 cells) cells quantified using Imaris spot detection tools from three replicates. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test. e Dot plots showing the average number of colocalized signals of IER3-AS1/IER3 (dsRNA) per cell in control (−D) and hnRNPK overexpressed (+D) cells. A total of 150 cells were manually counted from three independent replicates. Data are presented as mean values + / SEM. Statistical significance was calculated by Two-tailed unpaired t -test. For figures ( a , d and e ) * p ≤ 0.05; ** p ≤ 0.005; *** p ≤ 0.0005. f Model explaining the role of HnRNPK in the regulation of global dsRNA.

    Techniques Used: Immunofluorescence, Over Expression, Two Tailed Test, Western Blot

    human lung cancer cell line a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH human lung cancer cell line a549
    Human Lung Cancer Cell Line A549, supplied by CLS Cell Lines Service GmbH, 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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    human lung cancer cell line a549  (CLS Cell Lines Service GmbH)


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    CLS Cell Lines Service GmbH human lung cancer cell line a549
    Human Lung Cancer Cell Line A549, supplied by CLS Cell Lines Service GmbH, 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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    Structured Review

    CLS Cell Lines Service GmbH human lung cancer cell line a549
    Human Lung Cancer Cell Line A549, supplied by CLS Cell Lines Service GmbH, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human lung cancer cell line a549/product/CLS Cell Lines Service GmbH
    Average 93 stars, based on 1 article reviews
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    CLS Cell Lines Service GmbH a549 cells
    A549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH human lung adenocarcinoma cell line a549
    Human Lung Adenocarcinoma Cell Line A549, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH a 549 cells
    A 549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH cell lines a549
    Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. <t>A549</t> cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.
    Cell Lines A549, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH human non small cell lung carcinoma a549 cells
    Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from <t>A549</t> and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).
    Human Non Small Cell Lung Carcinoma A549 Cells, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH a549
    a , b Immunofluorescence images of <t>A549</t> cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.
    A549, supplied by CLS Cell Lines Service GmbH, 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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    CLS Cell Lines Service GmbH human lung cancer cell line a549
    a , b Immunofluorescence images of <t>A549</t> cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.
    Human Lung Cancer Cell Line A549, supplied by CLS Cell Lines Service GmbH, 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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    Image Search Results


    Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. A549 cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.

    Journal: Vaccines

    Article Title: The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling

    doi: 10.3390/vaccines11040768

    Figure Lengend Snippet: Effect of SARS-CoV-2 Spike 1 protein on epidermal growth factor receptor (EGFR), Ak strain transforming (AKT) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. A549 cells were treated or not with EGF (5 μg/mL) for 15 min or with full-length Spike 1 (2.5 μg/mL) or with the spike receptor-binding domain (RBD) (5 μg/mL) protein for 5 min then whole-cell extracts were subjected to western blot analysis for phosphoEGFR (pEGFR), phosphoAKT (pAKT), phosphor ERK1/2 (pERK1/2) and their respective total levels.

    Article Snippet: We have used commercially available cell lines A549 (human non-small cell lung cancer), HT-29 (human colorectal adenocarcinoma) and HeLa (human cervical cancer) obtained from Cell Line Service (CLS)-GmbH.

    Techniques: Activation Assay, Binding Assay, Western Blot

    Time-course accumulation of pEGFR and pERK1/2 in A549 cells treated with full length Spike 1 protein and its RBD. Cells were treated or not with 2.5 μg/mL full length Spike protein ( A ) or 5 μg/mL RBD ( B ) at different times as indicated (5, 15, 30 and 60 min) and the protein levels of pEGFR and pERK1/2 and their respective total levels were determined by western blot. The treatment with EGF (5 μg/mL) was carried out for 15 min.

    Journal: Vaccines

    Article Title: The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling

    doi: 10.3390/vaccines11040768

    Figure Lengend Snippet: Time-course accumulation of pEGFR and pERK1/2 in A549 cells treated with full length Spike 1 protein and its RBD. Cells were treated or not with 2.5 μg/mL full length Spike protein ( A ) or 5 μg/mL RBD ( B ) at different times as indicated (5, 15, 30 and 60 min) and the protein levels of pEGFR and pERK1/2 and their respective total levels were determined by western blot. The treatment with EGF (5 μg/mL) was carried out for 15 min.

    Article Snippet: We have used commercially available cell lines A549 (human non-small cell lung cancer), HT-29 (human colorectal adenocarcinoma) and HeLa (human cervical cancer) obtained from Cell Line Service (CLS)-GmbH.

    Techniques: Western Blot

    Blockade of EGFR inhibits SARS-CoV-2 spike 1-mediated activation of AKT in A549 cells. A549 were first pre-treated with AG1478 (10 μM) for 15 min prior to treatment with EGF ( A , B ), full-length spike 1 protein or spike RBD protein ( B ) as described in . Whole cell extracts were resolved on SDS-PAGE and protein levels were determined by western blot.

    Journal: Vaccines

    Article Title: The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling

    doi: 10.3390/vaccines11040768

    Figure Lengend Snippet: Blockade of EGFR inhibits SARS-CoV-2 spike 1-mediated activation of AKT in A549 cells. A549 were first pre-treated with AG1478 (10 μM) for 15 min prior to treatment with EGF ( A , B ), full-length spike 1 protein or spike RBD protein ( B ) as described in . Whole cell extracts were resolved on SDS-PAGE and protein levels were determined by western blot.

    Article Snippet: We have used commercially available cell lines A549 (human non-small cell lung cancer), HT-29 (human colorectal adenocarcinoma) and HeLa (human cervical cancer) obtained from Cell Line Service (CLS)-GmbH.

    Techniques: Activation Assay, SDS Page, Western Blot

    SARS-CoV-2 full length spike 1 protein and its RBD promote survivin expression in A549 cells. A549 cells were treated or not with EGF (5 μg/mL) ( A , B ), full-length spike 1 protein (2.5 μg/mL) ( A ) or spike RBD protein (5 μg/mL) ( B ) at different times (5, 15, 30 and 60 min) as indicated and the protein level of survivin was determined by Western blot. ( C ) A549 cells were first pre-treated or not with AG1478 (10 μM) for 15 min prior to treatment with EGF (5 μg/mL) or spike RBD protein (5 μg/mL) and the protein level of survivin was determined as described above.

    Journal: Vaccines

    Article Title: The SARS-CoV-2 Spike Protein Activates the Epidermal Growth Factor Receptor-Mediated Signaling

    doi: 10.3390/vaccines11040768

    Figure Lengend Snippet: SARS-CoV-2 full length spike 1 protein and its RBD promote survivin expression in A549 cells. A549 cells were treated or not with EGF (5 μg/mL) ( A , B ), full-length spike 1 protein (2.5 μg/mL) ( A ) or spike RBD protein (5 μg/mL) ( B ) at different times (5, 15, 30 and 60 min) as indicated and the protein level of survivin was determined by Western blot. ( C ) A549 cells were first pre-treated or not with AG1478 (10 μM) for 15 min prior to treatment with EGF (5 μg/mL) or spike RBD protein (5 μg/mL) and the protein level of survivin was determined as described above.

    Article Snippet: We have used commercially available cell lines A549 (human non-small cell lung cancer), HT-29 (human colorectal adenocarcinoma) and HeLa (human cervical cancer) obtained from Cell Line Service (CLS)-GmbH.

    Techniques: Expressing, Western Blot

    Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from A549 and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).

    Journal: Frontiers in Oncology

    Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

    doi: 10.3389/fonc.2022.1045521

    Figure Lengend Snippet: Human NSCLC tumor-derived cell line characterization. (A) Morphology of cells from A549 and primary cell lines grown in vitro for more than 10 passages. Scale bars = 400 µm. Cells have been stained with crystal violet dye. (B) The relative expression of stem cell- and EMT-related transcription factors as determined by RT-qPCR (in respect to GAPDH ). Data are presented as the mean ± SD (N = 2).

    Article Snippet: Human non-small cell lung carcinoma A549 cells were purchased from CLS (Cell Lines Service GmbH, Eppelheim, Germany).

    Techniques: Derivative Assay, In Vitro, Staining, Expressing, Quantitative RT-PCR

    Characterization of cell lines (continued). Relative expression of mesenchymal and epithelial markers cytokeratin (A) and vimentin (B) (proteins of intermediate filaments family) and activated protein kinases phospho-ERK and phospho-AKT (C) as determined by the Western blot method. Representative pictures from more than 3 independent experiments are shown. Coomassie-stained polyacrylamide gels serve as loading controls. Numbers indicate cell lines as in <xref ref-type= Figure 1 ; the letter A stands for the cell line A549. " width="100%" height="100%">

    Journal: Frontiers in Oncology

    Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

    doi: 10.3389/fonc.2022.1045521

    Figure Lengend Snippet: Characterization of cell lines (continued). Relative expression of mesenchymal and epithelial markers cytokeratin (A) and vimentin (B) (proteins of intermediate filaments family) and activated protein kinases phospho-ERK and phospho-AKT (C) as determined by the Western blot method. Representative pictures from more than 3 independent experiments are shown. Coomassie-stained polyacrylamide gels serve as loading controls. Numbers indicate cell lines as in Figure 1 ; the letter A stands for the cell line A549.

    Article Snippet: Human non-small cell lung carcinoma A549 cells were purchased from CLS (Cell Lines Service GmbH, Eppelheim, Germany).

    Techniques: Expressing, Western Blot, Staining

    Phosphorylation of ERK1/2 and AKT in lung cancer-derived cell lines after cisplatin treatment. (A) The dynamics of AKT and ERK phosphorylation in A549 cells. (B) Changes in AKT and ERK phosphorylation in lung tumor-derived cell lines after 6 hours of cisplatin treatment. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. The concentration of cisplatin was 90 µM.

    Journal: Frontiers in Oncology

    Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

    doi: 10.3389/fonc.2022.1045521

    Figure Lengend Snippet: Phosphorylation of ERK1/2 and AKT in lung cancer-derived cell lines after cisplatin treatment. (A) The dynamics of AKT and ERK phosphorylation in A549 cells. (B) Changes in AKT and ERK phosphorylation in lung tumor-derived cell lines after 6 hours of cisplatin treatment. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. The concentration of cisplatin was 90 µM.

    Article Snippet: Human non-small cell lung carcinoma A549 cells were purchased from CLS (Cell Lines Service GmbH, Eppelheim, Germany).

    Techniques: Derivative Assay, Western Blot, Staining, Concentration Assay

    Dependence of the interplay between ERK and AKT signaling pathways on extracellular contacts. (A) Focal adhesion kinase inhibitor PF573228 (+Fi; 10 µM) attenuates the MEK kinase inhibitor selumetinib-induced increase of AKT phosphorylation in control and cisplatin-treated lung cancer-derived cells, except A549 cells. (B) PF573228 prevents the AKT inhibitor capivasertib-induced increase in ERK phosphorylation in control and cisplatin-treated lung cancer-derived cells. (C) Cells grown in suspension (Susp) under agitation, in contrast to adherent cells (Adh), do not show alternative kinase activation after the treatment with inhibitors. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), +cis – cisplatin (90 µM), CAP – capivasertib (10 µM), AKTi – AKT inhibitor VIII (10 µM).

    Journal: Frontiers in Oncology

    Article Title: Crosstalk between protein kinases AKT and ERK1/2 in human lung tumor-derived cell models

    doi: 10.3389/fonc.2022.1045521

    Figure Lengend Snippet: Dependence of the interplay between ERK and AKT signaling pathways on extracellular contacts. (A) Focal adhesion kinase inhibitor PF573228 (+Fi; 10 µM) attenuates the MEK kinase inhibitor selumetinib-induced increase of AKT phosphorylation in control and cisplatin-treated lung cancer-derived cells, except A549 cells. (B) PF573228 prevents the AKT inhibitor capivasertib-induced increase in ERK phosphorylation in control and cisplatin-treated lung cancer-derived cells. (C) Cells grown in suspension (Susp) under agitation, in contrast to adherent cells (Adh), do not show alternative kinase activation after the treatment with inhibitors. Representative Western blots are shown. Coomassie-stained protein gels are presented as loading controls. 6-hour-long exposures to the drugs were used. DM – vehicle control (DMSO), SEL – selumetinib (10 µM), +cis – cisplatin (90 µM), CAP – capivasertib (10 µM), AKTi – AKT inhibitor VIII (10 µM).

    Article Snippet: Human non-small cell lung carcinoma A549 cells were purchased from CLS (Cell Lines Service GmbH, Eppelheim, Germany).

    Techniques: Derivative Assay, Activation Assay, Western Blot, Staining

    a , b Immunofluorescence images of A549 cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.

    Journal: Nature Communications

    Article Title: HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells

    doi: 10.1038/s41467-022-32537-0

    Figure Lengend Snippet: a , b Immunofluorescence images of A549 cells ( a ) and BT-549 ( b ) showing dsRNA-specific J2 signals in hnRNPK KD cells. The panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar is 50um. Graphs show the number of J2 signals in hnRNPK KD cells quantified using Imaris spot detection tools. For A549 cell line - control: n = 59, siRNA1: n = 64 and siRNA2: n = 73 cells and for BT-549: control: n = 25, siRNA1: n = 37 and siRNA2: n = 44 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. Significance was calculated by One-way ANOVA (ordinary multiple comparisons test). c , e RNAscope images of A549 cells ( c ) and BT-549 cells ( e ) showing IER3 (red) and IER3-AS1 (green) in control and hnRNPK KD cells. Panel to the right shows the magnified images of the merged panels indicated by white box. White arrows marks IER3-AS1 (green dots) and yellow arrows marks colocalized IER3/IER3-AS1 dsRNA (Yellow dots). Indicative scale bar on the images is 50 um. Bar graph shows the number of IER3-AS1 and IER3 RNA signals per cell in control and hnRNPK KD cells quantified using Imaris spot detection tools. For A549 - control: n = 101, siRNA1: n = 78 and siRNA2: n = 97 cells and for BT-549 - control: n = 132, siRNA1: n = 44 and siRNA2: n = F36 cells were counted from three independent experiments. Data are presented as mean values +/−SEM. The significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test). d Immunoblot showing the levels of HnRNPK and IER3 in hnRNPK KD A549 cells. GAPDH was used as an internal loading control. Similar results were observed in an independent biological replicate. Histograms showing the relative band intensities from the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates.

    Article Snippet: HEK293T, HeLa, A549 and BT-549 cells were obtained from CLS Cell Lines Service.

    Techniques: Immunofluorescence, Western Blot

    a Representative immunofluorescence images showing the effect of doxycycline (DOX) induced overexpression (−DOX/+DOX) of HnRNPK (green) on global dsRNA as detected using J2 antibody (red) in A549 cells. Panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar on the images is 50 um. Bar graph to the right shows the number of J2 signals in HnRNPK non-induced (−DOX (−D); n = 81 cells) and HnRNPK induced (DOX+(+D); n = 53 cells) cells from three replicates quantified as RNA spots per cell using Imaris spot detection tools. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-tailed unpaired t -test. b Immunoblot showing the levels of HnRNPK and IER3 in −DOX (−D) and +DOX (+D) A549 cells. GAPDH was used as an internal loading control. The similar results were observed in an independent experiment. Histograms showing the relative band intensities of the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates. c RNAscope images of DOX (−D/+D) induced HnRNPK cells showing IER3 (red) and IER3-AS1 (green) in control (−D) and hnRNPK overexpressed (+D) cells. Panel to the right showing magnified images of the merged panel indicated by white box. Yellow arrows depict colocalized IER3/IER3-AS1 dsRNA (yellow dots). Indicative scale bar on the images is 50 um. d Bar graph showing the number of IER3-AS1 and IER3 RNA signals per cell in control (−D; n = 80 cells) and hnRNPK overexpressed (+D; n = 121 cells) cells quantified using Imaris spot detection tools from three replicates. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test. e Dot plots showing the average number of colocalized signals of IER3-AS1/IER3 (dsRNA) per cell in control (−D) and hnRNPK overexpressed (+D) cells. A total of 150 cells were manually counted from three independent replicates. Data are presented as mean values + / SEM. Statistical significance was calculated by Two-tailed unpaired t -test. For figures ( a , d and e ) * p ≤ 0.05; ** p ≤ 0.005; *** p ≤ 0.0005. f Model explaining the role of HnRNPK in the regulation of global dsRNA.

    Journal: Nature Communications

    Article Title: HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells

    doi: 10.1038/s41467-022-32537-0

    Figure Lengend Snippet: a Representative immunofluorescence images showing the effect of doxycycline (DOX) induced overexpression (−DOX/+DOX) of HnRNPK (green) on global dsRNA as detected using J2 antibody (red) in A549 cells. Panel to the right shows the merged images. Magnified J2 signals (puncta) are shown in an inset at the upper right corner indicated by white box. Indicative scale bar on the images is 50 um. Bar graph to the right shows the number of J2 signals in HnRNPK non-induced (−DOX (−D); n = 81 cells) and HnRNPK induced (DOX+(+D); n = 53 cells) cells from three replicates quantified as RNA spots per cell using Imaris spot detection tools. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-tailed unpaired t -test. b Immunoblot showing the levels of HnRNPK and IER3 in −DOX (−D) and +DOX (+D) A549 cells. GAPDH was used as an internal loading control. The similar results were observed in an independent experiment. Histograms showing the relative band intensities of the western blot are shown left side. The p values were calculated using two-sided Student’s t -test and data are presented ± SD from two independent replicates. c RNAscope images of DOX (−D/+D) induced HnRNPK cells showing IER3 (red) and IER3-AS1 (green) in control (−D) and hnRNPK overexpressed (+D) cells. Panel to the right showing magnified images of the merged panel indicated by white box. Yellow arrows depict colocalized IER3/IER3-AS1 dsRNA (yellow dots). Indicative scale bar on the images is 50 um. d Bar graph showing the number of IER3-AS1 and IER3 RNA signals per cell in control (−D; n = 80 cells) and hnRNPK overexpressed (+D; n = 121 cells) cells quantified using Imaris spot detection tools from three replicates. Data are presented as mean values +/−SEM. Statistical significance was calculated by Two-way ANOVA (Sidak’s multiple comparisons test. e Dot plots showing the average number of colocalized signals of IER3-AS1/IER3 (dsRNA) per cell in control (−D) and hnRNPK overexpressed (+D) cells. A total of 150 cells were manually counted from three independent replicates. Data are presented as mean values + / SEM. Statistical significance was calculated by Two-tailed unpaired t -test. For figures ( a , d and e ) * p ≤ 0.05; ** p ≤ 0.005; *** p ≤ 0.0005. f Model explaining the role of HnRNPK in the regulation of global dsRNA.

    Article Snippet: HEK293T, HeLa, A549 and BT-549 cells were obtained from CLS Cell Lines Service.

    Techniques: Immunofluorescence, Over Expression, Two Tailed Test, Western Blot