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wild type wt human lung epithelial a549 cells  (ATCC)


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    ATCC wild type wt human lung epithelial a549 cells
    C3 deficiency alters gene expression in <t>human</t> <t>lung</t> <t>epithelial</t> <t>A549</t> cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1
    Wild Type Wt Human Lung Epithelial A549 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 35523 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wild type wt human lung epithelial a549 cells/product/ATCC
    Average 99 stars, based on 35523 article reviews
    wild type wt human lung epithelial a549 cells - by Bioz Stars, 2026-02
    99/100 stars

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    1) Product Images from "Intracellular C3 regulates the immune response to infection via NF-κB signaling"

    Article Title: Intracellular C3 regulates the immune response to infection via NF-κB signaling

    Journal: Cellular and Molecular Life Sciences: CMLS

    doi: 10.1007/s00018-025-05975-4

    C3 deficiency alters gene expression in human lung epithelial A549 cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1
    Figure Legend Snippet: C3 deficiency alters gene expression in human lung epithelial A549 cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1

    Techniques Used: Gene Expression, Expressing, Western Blot, Clone Assay, RNA Sequencing

    Cytokine secretion from A549 cells is modulated by the presence of C3. XL cytokine array analysis of supernatants from WT and C3 KO A549 cells infected with M. catarrhalis shows lower secretion from C3 KO cells. Reference spots and negative controls are indicated by + and –, respectively ( A ) with quantification in ( B ), and black arrows indicating IL-8 and MCP-1, as differentially secreted cytokines. Bio-Plex Pro human cytokine 27-plex assay with supernatants from WT, C3 KO and C3 ΔATG1 cells infected with M. catarrhalis ( C ) or Y. enterocolitica ( D ) shows lower secretion from C3 KO cells that is rescued in C3 ΔATG1. Bars represent the average of three biological repetitions indicated with symbols. Two-way ANOVA with Tukey´s multiple comparison test, matched measures within the experiment, * p < 0.05, **** p < 0.0001
    Figure Legend Snippet: Cytokine secretion from A549 cells is modulated by the presence of C3. XL cytokine array analysis of supernatants from WT and C3 KO A549 cells infected with M. catarrhalis shows lower secretion from C3 KO cells. Reference spots and negative controls are indicated by + and –, respectively ( A ) with quantification in ( B ), and black arrows indicating IL-8 and MCP-1, as differentially secreted cytokines. Bio-Plex Pro human cytokine 27-plex assay with supernatants from WT, C3 KO and C3 ΔATG1 cells infected with M. catarrhalis ( C ) or Y. enterocolitica ( D ) shows lower secretion from C3 KO cells that is rescued in C3 ΔATG1. Bars represent the average of three biological repetitions indicated with symbols. Two-way ANOVA with Tukey´s multiple comparison test, matched measures within the experiment, * p < 0.05, **** p < 0.0001

    Techniques Used: Infection, Plex Assay, Comparison

    Cytosolic C3 contributes to cytokine release after stimulation of different receptors. ( A - F ) Levels of cytokine secretion measured by ELISA in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells after infection with bacteria are reduced in cells lacking C3. IL-8 secretion after infection with M. catarrhalis ( A ), IL-6 after infection Y. enterocolitica ( B ), RANTES secretion after infection with M. catarrhalis ( C ), RANTES secretion after infection with Y. enterocolitica ( D ), MCP-1 secretion after infection with M. catarrhalis ( E ), and MCP-1 secretion after infection with Y. enterocolitica ( F ). ( G - M ) Levels of cytokine secretion in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells are reduced in cells lacking C3 after stimulation of TLR or TNFR, but not after IL-1R stimulation. IL-8 secretion after treatment with Pam3CSK4 ( G ), IL-6 secretion after treatment with TNF ( H ), sICAM-1 release after treatment with Pam3CSK4 ( I ), sICAM-1 release after TNF treatment ( J ), IL-8 secretion after poly(I: C) treatment ( K ), MCP-1 secretion after poly(I: C) treatment ( L ), and IL-8 secretion after IL-1β treatment ( M ). Results are shown as mean ± SD of independent experiments. Each dot represents an average of at least three clones in one biological repeat. The cytokine secretion levels of untreated cells were subtracted from the values of treated cells. One-way ANOVA with Tukey´s multiple comparison test, matched measures within experiment for C-F and I-M. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant
    Figure Legend Snippet: Cytosolic C3 contributes to cytokine release after stimulation of different receptors. ( A - F ) Levels of cytokine secretion measured by ELISA in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells after infection with bacteria are reduced in cells lacking C3. IL-8 secretion after infection with M. catarrhalis ( A ), IL-6 after infection Y. enterocolitica ( B ), RANTES secretion after infection with M. catarrhalis ( C ), RANTES secretion after infection with Y. enterocolitica ( D ), MCP-1 secretion after infection with M. catarrhalis ( E ), and MCP-1 secretion after infection with Y. enterocolitica ( F ). ( G - M ) Levels of cytokine secretion in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells are reduced in cells lacking C3 after stimulation of TLR or TNFR, but not after IL-1R stimulation. IL-8 secretion after treatment with Pam3CSK4 ( G ), IL-6 secretion after treatment with TNF ( H ), sICAM-1 release after treatment with Pam3CSK4 ( I ), sICAM-1 release after TNF treatment ( J ), IL-8 secretion after poly(I: C) treatment ( K ), MCP-1 secretion after poly(I: C) treatment ( L ), and IL-8 secretion after IL-1β treatment ( M ). Results are shown as mean ± SD of independent experiments. Each dot represents an average of at least three clones in one biological repeat. The cytokine secretion levels of untreated cells were subtracted from the values of treated cells. One-way ANOVA with Tukey´s multiple comparison test, matched measures within experiment for C-F and I-M. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Bacteria, Clone Assay, Comparison

    Cytokine gene transcription and NF-κB pathway activation are dependent on cytosolic C3. mRNA levels for IL-6 are decreased in C3 KO A549 cells compared to WT and C3 ΔATG1 cells after infection with Y. enterocolitica , or treatment with TNF ( A ). Similar is observed for mRNA levels for IL-8 after infection with M. catarrhalis or Y. enterocolitica , however treatment with TNF or Pam3CSK4 does not reach significance ( B ). NF-κB activation assessed using luciferase reporter assay is decreased in C3 KO compared to WT and C3 ΔATG1 cells after treatment with Pam3CSK4 ( C ) or infection with M. catarrhalis ( D ). Representative Western blot showing NF-κB (p65 subunit) translocation to the nucleus ( E ) with quantification in ( F ). C3 KO cells are characterized by diminished translocation of p65 to the nucleus compared to WT and C3 ΔATG1;- and + indicate no treatment and treatment with Pam3CSK4, respectively. A similar result was obtained upon infection with M. catarrhalis , a representative blot of nuclear fractions shown in ( G ) with quantification in ( H ). Representative Western blot analysis showing decreased phosphorylation of IKK in lysates from C3 KO cells compared to WT; - and + indicate no treatment and treatment with Pam3CSK4 ( I ) or M. catarrhalis ( K ) with quantification in ( J ) and ( L ), respectively. Each dot represents one biological repeat for one clone ( A , B ) or an average of two clones in one biological repeat ( C - L ). One-way ( C , D ) or Two-way ( A , B , F , H , J , L ) ANOVA with Tukey´s multiple comparison test, matched measures within the experiment. *p< 0.05, **p < 0.01, ***p < 0.001, ns: not significant
    Figure Legend Snippet: Cytokine gene transcription and NF-κB pathway activation are dependent on cytosolic C3. mRNA levels for IL-6 are decreased in C3 KO A549 cells compared to WT and C3 ΔATG1 cells after infection with Y. enterocolitica , or treatment with TNF ( A ). Similar is observed for mRNA levels for IL-8 after infection with M. catarrhalis or Y. enterocolitica , however treatment with TNF or Pam3CSK4 does not reach significance ( B ). NF-κB activation assessed using luciferase reporter assay is decreased in C3 KO compared to WT and C3 ΔATG1 cells after treatment with Pam3CSK4 ( C ) or infection with M. catarrhalis ( D ). Representative Western blot showing NF-κB (p65 subunit) translocation to the nucleus ( E ) with quantification in ( F ). C3 KO cells are characterized by diminished translocation of p65 to the nucleus compared to WT and C3 ΔATG1;- and + indicate no treatment and treatment with Pam3CSK4, respectively. A similar result was obtained upon infection with M. catarrhalis , a representative blot of nuclear fractions shown in ( G ) with quantification in ( H ). Representative Western blot analysis showing decreased phosphorylation of IKK in lysates from C3 KO cells compared to WT; - and + indicate no treatment and treatment with Pam3CSK4 ( I ) or M. catarrhalis ( K ) with quantification in ( J ) and ( L ), respectively. Each dot represents one biological repeat for one clone ( A , B ) or an average of two clones in one biological repeat ( C - L ). One-way ( C , D ) or Two-way ( A , B , F , H , J , L ) ANOVA with Tukey´s multiple comparison test, matched measures within the experiment. *p< 0.05, **p < 0.01, ***p < 0.001, ns: not significant

    Techniques Used: Activation Assay, Infection, Luciferase, Reporter Assay, Western Blot, Translocation Assay, Phospho-proteomics, Clone Assay, Comparison



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    wild type wt human lung epithelial a549 cells  (ATCC)
    99
    ATCC wild type wt human lung epithelial a549 cells
    C3 deficiency alters gene expression in <t>human</t> <t>lung</t> <t>epithelial</t> <t>A549</t> cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1
    Wild Type Wt Human Lung Epithelial A549 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wild type wt human lung epithelial a549 cells/product/ATCC
    Average 99 stars, based on 1 article reviews
    wild type wt human lung epithelial a549 cells - by Bioz Stars, 2026-02
    99/100 stars
    		  Buy from Supplier

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    C3 deficiency alters gene expression in human lung epithelial A549 cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: Intracellular C3 regulates the immune response to infection via NF-κB signaling

    doi: 10.1007/s00018-025-05975-4

    Figure Lengend Snippet: C3 deficiency alters gene expression in human lung epithelial A549 cell line, which is rescued by cytosolic C3. A ) Three variants of A549 cells expressing C3 (WT), entirely lacking C3 (C3 KO) or expressing only cytosolic C3 (C3 ΔATG1) were used in this study. Western blotting analysis of cell lysates showing the presence of pro-C3 along with mature alpha and beta chains in WT cells ( B ). No C3 was detected in C3 KO cells while C3 ΔATG1 clones expressed only pro-C3 ( C ). D ) While WT and C3 ΔATG1 cells showed similar gene expression, C3 KO cells had strongly altered gene expression compared to WT and C3 ΔATG1 (padj < 0.05). E ) Heat map representation of the expression patterns in WT, C3 KO and C3 ΔATG1 clones (4 clones for each genotype) and F ) Principal component (PC) analysis of RNA-seq data for WT, C3 KO and C3 ΔATG1 clones showed that C3 KO clones cluster separately from WT and C3 ΔATG1 clones. G ) Venn diagram of DEGs across different genotypes. Summary of KEGG enrichment analysis showing pathways with >2 differentially expressed genes between WT and C3 KO ( H ) and between C3 KO and C3 ΔATG1 ( I ). For analysis of differentially regulated genes, we used the following criteria: pair-wise analysis with padj≤ 0.05 & |log2(foldchange)| ≥ 1

    Article Snippet: The wild-type (WT) human lung epithelial A549 cells were from the American Type Culture Collection (ATCC).

    Techniques: Gene Expression, Expressing, Western Blot, Clone Assay, RNA Sequencing

    Cytokine secretion from A549 cells is modulated by the presence of C3. XL cytokine array analysis of supernatants from WT and C3 KO A549 cells infected with M. catarrhalis shows lower secretion from C3 KO cells. Reference spots and negative controls are indicated by + and –, respectively ( A ) with quantification in ( B ), and black arrows indicating IL-8 and MCP-1, as differentially secreted cytokines. Bio-Plex Pro human cytokine 27-plex assay with supernatants from WT, C3 KO and C3 ΔATG1 cells infected with M. catarrhalis ( C ) or Y. enterocolitica ( D ) shows lower secretion from C3 KO cells that is rescued in C3 ΔATG1. Bars represent the average of three biological repetitions indicated with symbols. Two-way ANOVA with Tukey´s multiple comparison test, matched measures within the experiment, * p < 0.05, **** p < 0.0001

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: Intracellular C3 regulates the immune response to infection via NF-κB signaling

    doi: 10.1007/s00018-025-05975-4

    Figure Lengend Snippet: Cytokine secretion from A549 cells is modulated by the presence of C3. XL cytokine array analysis of supernatants from WT and C3 KO A549 cells infected with M. catarrhalis shows lower secretion from C3 KO cells. Reference spots and negative controls are indicated by + and –, respectively ( A ) with quantification in ( B ), and black arrows indicating IL-8 and MCP-1, as differentially secreted cytokines. Bio-Plex Pro human cytokine 27-plex assay with supernatants from WT, C3 KO and C3 ΔATG1 cells infected with M. catarrhalis ( C ) or Y. enterocolitica ( D ) shows lower secretion from C3 KO cells that is rescued in C3 ΔATG1. Bars represent the average of three biological repetitions indicated with symbols. Two-way ANOVA with Tukey´s multiple comparison test, matched measures within the experiment, * p < 0.05, **** p < 0.0001

    Article Snippet: The wild-type (WT) human lung epithelial A549 cells were from the American Type Culture Collection (ATCC).

    Techniques: Infection, Plex Assay, Comparison

    Cytosolic C3 contributes to cytokine release after stimulation of different receptors. ( A - F ) Levels of cytokine secretion measured by ELISA in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells after infection with bacteria are reduced in cells lacking C3. IL-8 secretion after infection with M. catarrhalis ( A ), IL-6 after infection Y. enterocolitica ( B ), RANTES secretion after infection with M. catarrhalis ( C ), RANTES secretion after infection with Y. enterocolitica ( D ), MCP-1 secretion after infection with M. catarrhalis ( E ), and MCP-1 secretion after infection with Y. enterocolitica ( F ). ( G - M ) Levels of cytokine secretion in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells are reduced in cells lacking C3 after stimulation of TLR or TNFR, but not after IL-1R stimulation. IL-8 secretion after treatment with Pam3CSK4 ( G ), IL-6 secretion after treatment with TNF ( H ), sICAM-1 release after treatment with Pam3CSK4 ( I ), sICAM-1 release after TNF treatment ( J ), IL-8 secretion after poly(I: C) treatment ( K ), MCP-1 secretion after poly(I: C) treatment ( L ), and IL-8 secretion after IL-1β treatment ( M ). Results are shown as mean ± SD of independent experiments. Each dot represents an average of at least three clones in one biological repeat. The cytokine secretion levels of untreated cells were subtracted from the values of treated cells. One-way ANOVA with Tukey´s multiple comparison test, matched measures within experiment for C-F and I-M. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: Intracellular C3 regulates the immune response to infection via NF-κB signaling

    doi: 10.1007/s00018-025-05975-4

    Figure Lengend Snippet: Cytosolic C3 contributes to cytokine release after stimulation of different receptors. ( A - F ) Levels of cytokine secretion measured by ELISA in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells after infection with bacteria are reduced in cells lacking C3. IL-8 secretion after infection with M. catarrhalis ( A ), IL-6 after infection Y. enterocolitica ( B ), RANTES secretion after infection with M. catarrhalis ( C ), RANTES secretion after infection with Y. enterocolitica ( D ), MCP-1 secretion after infection with M. catarrhalis ( E ), and MCP-1 secretion after infection with Y. enterocolitica ( F ). ( G - M ) Levels of cytokine secretion in supernatants from WT, C3 KO and C3 ΔATG1 A549 cells are reduced in cells lacking C3 after stimulation of TLR or TNFR, but not after IL-1R stimulation. IL-8 secretion after treatment with Pam3CSK4 ( G ), IL-6 secretion after treatment with TNF ( H ), sICAM-1 release after treatment with Pam3CSK4 ( I ), sICAM-1 release after TNF treatment ( J ), IL-8 secretion after poly(I: C) treatment ( K ), MCP-1 secretion after poly(I: C) treatment ( L ), and IL-8 secretion after IL-1β treatment ( M ). Results are shown as mean ± SD of independent experiments. Each dot represents an average of at least three clones in one biological repeat. The cytokine secretion levels of untreated cells were subtracted from the values of treated cells. One-way ANOVA with Tukey´s multiple comparison test, matched measures within experiment for C-F and I-M. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant

    Article Snippet: The wild-type (WT) human lung epithelial A549 cells were from the American Type Culture Collection (ATCC).

    Techniques: Enzyme-linked Immunosorbent Assay, Infection, Bacteria, Clone Assay, Comparison

    Cytokine gene transcription and NF-κB pathway activation are dependent on cytosolic C3. mRNA levels for IL-6 are decreased in C3 KO A549 cells compared to WT and C3 ΔATG1 cells after infection with Y. enterocolitica , or treatment with TNF ( A ). Similar is observed for mRNA levels for IL-8 after infection with M. catarrhalis or Y. enterocolitica , however treatment with TNF or Pam3CSK4 does not reach significance ( B ). NF-κB activation assessed using luciferase reporter assay is decreased in C3 KO compared to WT and C3 ΔATG1 cells after treatment with Pam3CSK4 ( C ) or infection with M. catarrhalis ( D ). Representative Western blot showing NF-κB (p65 subunit) translocation to the nucleus ( E ) with quantification in ( F ). C3 KO cells are characterized by diminished translocation of p65 to the nucleus compared to WT and C3 ΔATG1;- and + indicate no treatment and treatment with Pam3CSK4, respectively. A similar result was obtained upon infection with M. catarrhalis , a representative blot of nuclear fractions shown in ( G ) with quantification in ( H ). Representative Western blot analysis showing decreased phosphorylation of IKK in lysates from C3 KO cells compared to WT; - and + indicate no treatment and treatment with Pam3CSK4 ( I ) or M. catarrhalis ( K ) with quantification in ( J ) and ( L ), respectively. Each dot represents one biological repeat for one clone ( A , B ) or an average of two clones in one biological repeat ( C - L ). One-way ( C , D ) or Two-way ( A , B , F , H , J , L ) ANOVA with Tukey´s multiple comparison test, matched measures within the experiment. *p< 0.05, **p < 0.01, ***p < 0.001, ns: not significant

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: Intracellular C3 regulates the immune response to infection via NF-κB signaling

    doi: 10.1007/s00018-025-05975-4

    Figure Lengend Snippet: Cytokine gene transcription and NF-κB pathway activation are dependent on cytosolic C3. mRNA levels for IL-6 are decreased in C3 KO A549 cells compared to WT and C3 ΔATG1 cells after infection with Y. enterocolitica , or treatment with TNF ( A ). Similar is observed for mRNA levels for IL-8 after infection with M. catarrhalis or Y. enterocolitica , however treatment with TNF or Pam3CSK4 does not reach significance ( B ). NF-κB activation assessed using luciferase reporter assay is decreased in C3 KO compared to WT and C3 ΔATG1 cells after treatment with Pam3CSK4 ( C ) or infection with M. catarrhalis ( D ). Representative Western blot showing NF-κB (p65 subunit) translocation to the nucleus ( E ) with quantification in ( F ). C3 KO cells are characterized by diminished translocation of p65 to the nucleus compared to WT and C3 ΔATG1;- and + indicate no treatment and treatment with Pam3CSK4, respectively. A similar result was obtained upon infection with M. catarrhalis , a representative blot of nuclear fractions shown in ( G ) with quantification in ( H ). Representative Western blot analysis showing decreased phosphorylation of IKK in lysates from C3 KO cells compared to WT; - and + indicate no treatment and treatment with Pam3CSK4 ( I ) or M. catarrhalis ( K ) with quantification in ( J ) and ( L ), respectively. Each dot represents one biological repeat for one clone ( A , B ) or an average of two clones in one biological repeat ( C - L ). One-way ( C , D ) or Two-way ( A , B , F , H , J , L ) ANOVA with Tukey´s multiple comparison test, matched measures within the experiment. *p< 0.05, **p < 0.01, ***p < 0.001, ns: not significant

    Article Snippet: The wild-type (WT) human lung epithelial A549 cells were from the American Type Culture Collection (ATCC).

    Techniques: Activation Assay, Infection, Luciferase, Reporter Assay, Western Blot, Translocation Assay, Phospho-proteomics, Clone Assay, Comparison