Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Cell lines used for this study

Article Snippet: THP1 , ATCC (TIB 202) , Human monocytes from acute monocytic leukemia , RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine.

Techniques:

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Responses of 19 mammalian cell lines to infection with H. pylori strain P1

Article Snippet: THP1 , ATCC (TIB 202) , Human monocytes from acute monocytic leukemia , RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine.

Techniques: Infection

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Chemokine secretion from H. pylori-infected human and murine cell lines. Cells were infected for 20 h with the H. pylori wild-type strain (P1wt) or the type IV secretion-defective knockout mutant P1ΔvirB11. IL-8 or Mip-2 secreted into the medium was determined by ELISA. The amounts of chemokine secretion are presented as percentages relative to the amount during wild-type infection, except for GLC4, L929, SR4987, and SYF+src cells, from which no secretion was detected at all. For most cell lines, chemokine secretion was dependent on the function of the TFSS. In addition, HeLa and THP1 cells showed some IL-8 secretion upon infection withP1ΔvirB11. The Mann-Whitney test indicated that this low level of IL-8 secretion was significantly higher than that in noninfected cells (P < 0.05, asterisks). Infection of the mouse cell line J774.A with P1ΔvirB11 induced a slightly higher Mip-2 secretion level than infection with the wild-type strain. Error bars indicate standard deviations.

Article Snippet: THP1 , ATCC (TIB 202) , Human monocytes from acute monocytic leukemia , RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine.

Techniques: Infection, Knock-Out, Mutagenesis, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Phenotypes of H. pylori-infected host cells. Infected cells were analyzed by phase-contrast microscopy. Only AGS cells showed the clearly defined hummingbird phenotype (upper panels). The hummingbird phenotype requires CagA, as it was not observed after infection with P1ΔcagA (not shown). THP1 cells formed aggregates after infection with the wild-type strain (middle panels). Aggregation is independent of CagA but requires the TFSS, as it was induced upon infection with the cagA but not the virB11 knockout mutant (not shown). Kato3 cells and all other cell lines tested did not reveal any obvious change in phenotype upon infection (lower panels). Bar, 100 μm.

Article Snippet: THP1 , ATCC (TIB 202) , Human monocytes from acute monocytic leukemia , RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine.

Techniques: Infection, Microscopy, Knock-Out, Mutagenesis

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Model of the roles of host cell factors required for the cellular responses mediated by the H. pylori TFSS. H. pylori translocates CagA, which is phosphorylated by Src family kinases (SFK). Tyrosine-phosphorylated CagA has been shown to interact with Csk and SHP-2, to inactivate Src family kinases, and to induce tyrosine dephosphorylation of cortactin and ezrin and rearrangements of the actin cytoskeleton. The TFSS also translocates peptidoglycan (PG), which activates the intracellular Nod1 receptor, activates the transcription factor NF-κB, and stimulates the expression and secretion of IL-8 and the upregulation of ICAM-1 on the cell surface. (A) Host cell factors appear to be required for CagA and peptidoglycan translocation. These factors could be inducers or cellular receptors of the TFSS. (B) Processing of CagA only occurs in some cell lines, indicating that specific cellular proteases are involved. (C) The hummingbird phenotype is only observed with AGS cells. Thus, some unknown host cell factors are apparently required for its induction. (D) HeLa and THP1 cells secrete some IL-8 in a TFSS-independent manner, indicating that another receptor can detect H. pylori in these cells.

Article Snippet: THP1 , ATCC (TIB 202) , Human monocytes from acute monocytic leukemia , RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine.

Techniques: De-Phosphorylation Assay, Expressing, Translocation Assay

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Cell lines used for this study

Article Snippet: Cells were grown in the indicated media and subcultured every 2 to 3 days. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Cell line Reference or source a (no.) Cell type Growth medium b 293T ATCC (CRL 11268) Human embryonal kidney DMEM (Gibco BRL), 10% fetal bovine serum (FBS; Gibco BRL), 2 mM l -glutamine, 1 mM sodium pyruvate AGS ATCC (CRL 1739) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine CHO K1 ATCC (CCL 61) Chinese hamster ovary MEM alpha (Gibco BRL), 10% FBS Cos1 ATCC (CRL 1650) Green African monkey kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine GLC4 42 Human lung carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Hec1.b ATCC (HTP 113) Human endometrial adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine Hela ATCC (CCL 2) Human cervix epithelial carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HepG2 ATCC (HB 8065) Human hepatocellular carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HL ECACC (96121720) Human lung, fibroblast-like RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HT29 ATCC (HTB 38) Human colon adenocarcinoma Mc Coy's 5A (Gibco BRL), 10% FBS J774.A DSMZ (ACC 170) Mouse monocytic macrophages RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Kato3 ATCC (HTB 103) Human gastic carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine L929 ATCC (CCL 1) Mouse fibroblasts RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MDCK ATCC (CCL 34) Canine kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MKN-28 27 Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine MKN-45 DSMZ (ACC 409) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine SR4987 DSMZ (ACC 323) Mouse stromal fibroblast-like Mc Coy's 5A (Gibco BRL), 10% FBS SYF+src ATCC (CRL 2498) Mouse fibroblasts overexpressing Src DMEM (Gibco BRL), 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate THP1 ATCC (TIB 202) Human monocytes from acute monocytic leukemia RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Open in a separate window a ATCC, American Type Culture Collection ( www.atcc.org ); DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH ( www.dsmz.de ); ECACC, European Collection of Cell Cultures ( www.ecacc.org.uk ). b DMEM, Dulbecco's modified Eagle's medium; MEM, minimum essential medium.

Techniques:

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Responses of 19 mammalian cell lines to infection with H. pylori strain P1

Article Snippet: Cells were grown in the indicated media and subcultured every 2 to 3 days. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Cell line Reference or source a (no.) Cell type Growth medium b 293T ATCC (CRL 11268) Human embryonal kidney DMEM (Gibco BRL), 10% fetal bovine serum (FBS; Gibco BRL), 2 mM l -glutamine, 1 mM sodium pyruvate AGS ATCC (CRL 1739) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine CHO K1 ATCC (CCL 61) Chinese hamster ovary MEM alpha (Gibco BRL), 10% FBS Cos1 ATCC (CRL 1650) Green African monkey kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine GLC4 42 Human lung carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Hec1.b ATCC (HTP 113) Human endometrial adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine Hela ATCC (CCL 2) Human cervix epithelial carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HepG2 ATCC (HB 8065) Human hepatocellular carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HL ECACC (96121720) Human lung, fibroblast-like RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HT29 ATCC (HTB 38) Human colon adenocarcinoma Mc Coy's 5A (Gibco BRL), 10% FBS J774.A DSMZ (ACC 170) Mouse monocytic macrophages RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Kato3 ATCC (HTB 103) Human gastic carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine L929 ATCC (CCL 1) Mouse fibroblasts RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MDCK ATCC (CCL 34) Canine kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MKN-28 27 Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine MKN-45 DSMZ (ACC 409) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine SR4987 DSMZ (ACC 323) Mouse stromal fibroblast-like Mc Coy's 5A (Gibco BRL), 10% FBS SYF+src ATCC (CRL 2498) Mouse fibroblasts overexpressing Src DMEM (Gibco BRL), 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate THP1 ATCC (TIB 202) Human monocytes from acute monocytic leukemia RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Open in a separate window a ATCC, American Type Culture Collection ( www.atcc.org ); DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH ( www.dsmz.de ); ECACC, European Collection of Cell Cultures ( www.ecacc.org.uk ). b DMEM, Dulbecco's modified Eagle's medium; MEM, minimum essential medium.

Techniques: Infection

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Chemokine secretion from H. pylori-infected human and murine cell lines. Cells were infected for 20 h with the H. pylori wild-type strain (P1wt) or the type IV secretion-defective knockout mutant P1ΔvirB11. IL-8 or Mip-2 secreted into the medium was determined by ELISA. The amounts of chemokine secretion are presented as percentages relative to the amount during wild-type infection, except for GLC4, L929, SR4987, and SYF+src cells, from which no secretion was detected at all. For most cell lines, chemokine secretion was dependent on the function of the TFSS. In addition, HeLa and THP1 cells showed some IL-8 secretion upon infection withP1ΔvirB11. The Mann-Whitney test indicated that this low level of IL-8 secretion was significantly higher than that in noninfected cells (P < 0.05, asterisks). Infection of the mouse cell line J774.A with P1ΔvirB11 induced a slightly higher Mip-2 secretion level than infection with the wild-type strain. Error bars indicate standard deviations.

Article Snippet: Cells were grown in the indicated media and subcultured every 2 to 3 days. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Cell line Reference or source a (no.) Cell type Growth medium b 293T ATCC (CRL 11268) Human embryonal kidney DMEM (Gibco BRL), 10% fetal bovine serum (FBS; Gibco BRL), 2 mM l -glutamine, 1 mM sodium pyruvate AGS ATCC (CRL 1739) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine CHO K1 ATCC (CCL 61) Chinese hamster ovary MEM alpha (Gibco BRL), 10% FBS Cos1 ATCC (CRL 1650) Green African monkey kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine GLC4 42 Human lung carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Hec1.b ATCC (HTP 113) Human endometrial adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine Hela ATCC (CCL 2) Human cervix epithelial carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HepG2 ATCC (HB 8065) Human hepatocellular carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HL ECACC (96121720) Human lung, fibroblast-like RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HT29 ATCC (HTB 38) Human colon adenocarcinoma Mc Coy's 5A (Gibco BRL), 10% FBS J774.A DSMZ (ACC 170) Mouse monocytic macrophages RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Kato3 ATCC (HTB 103) Human gastic carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine L929 ATCC (CCL 1) Mouse fibroblasts RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MDCK ATCC (CCL 34) Canine kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MKN-28 27 Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine MKN-45 DSMZ (ACC 409) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine SR4987 DSMZ (ACC 323) Mouse stromal fibroblast-like Mc Coy's 5A (Gibco BRL), 10% FBS SYF+src ATCC (CRL 2498) Mouse fibroblasts overexpressing Src DMEM (Gibco BRL), 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate THP1 ATCC (TIB 202) Human monocytes from acute monocytic leukemia RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Open in a separate window a ATCC, American Type Culture Collection ( www.atcc.org ); DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH ( www.dsmz.de ); ECACC, European Collection of Cell Cultures ( www.ecacc.org.uk ). b DMEM, Dulbecco's modified Eagle's medium; MEM, minimum essential medium.

Techniques: Infection, Knock-Out, Mutagenesis, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Phenotypes of H. pylori-infected host cells. Infected cells were analyzed by phase-contrast microscopy. Only AGS cells showed the clearly defined hummingbird phenotype (upper panels). The hummingbird phenotype requires CagA, as it was not observed after infection with P1ΔcagA (not shown). THP1 cells formed aggregates after infection with the wild-type strain (middle panels). Aggregation is independent of CagA but requires the TFSS, as it was induced upon infection with the cagA but not the virB11 knockout mutant (not shown). Kato3 cells and all other cell lines tested did not reveal any obvious change in phenotype upon infection (lower panels). Bar, 100 μm.

Article Snippet: Cells were grown in the indicated media and subcultured every 2 to 3 days. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Cell line Reference or source a (no.) Cell type Growth medium b 293T ATCC (CRL 11268) Human embryonal kidney DMEM (Gibco BRL), 10% fetal bovine serum (FBS; Gibco BRL), 2 mM l -glutamine, 1 mM sodium pyruvate AGS ATCC (CRL 1739) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine CHO K1 ATCC (CCL 61) Chinese hamster ovary MEM alpha (Gibco BRL), 10% FBS Cos1 ATCC (CRL 1650) Green African monkey kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine GLC4 42 Human lung carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Hec1.b ATCC (HTP 113) Human endometrial adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine Hela ATCC (CCL 2) Human cervix epithelial carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HepG2 ATCC (HB 8065) Human hepatocellular carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HL ECACC (96121720) Human lung, fibroblast-like RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HT29 ATCC (HTB 38) Human colon adenocarcinoma Mc Coy's 5A (Gibco BRL), 10% FBS J774.A DSMZ (ACC 170) Mouse monocytic macrophages RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Kato3 ATCC (HTB 103) Human gastic carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine L929 ATCC (CCL 1) Mouse fibroblasts RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MDCK ATCC (CCL 34) Canine kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MKN-28 27 Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine MKN-45 DSMZ (ACC 409) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine SR4987 DSMZ (ACC 323) Mouse stromal fibroblast-like Mc Coy's 5A (Gibco BRL), 10% FBS SYF+src ATCC (CRL 2498) Mouse fibroblasts overexpressing Src DMEM (Gibco BRL), 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate THP1 ATCC (TIB 202) Human monocytes from acute monocytic leukemia RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Open in a separate window a ATCC, American Type Culture Collection ( www.atcc.org ); DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH ( www.dsmz.de ); ECACC, European Collection of Cell Cultures ( www.ecacc.org.uk ). b DMEM, Dulbecco's modified Eagle's medium; MEM, minimum essential medium.

Techniques: Infection, Microscopy, Knock-Out, Mutagenesis

Journal:

Article Title: Analysis of Cell Type-Specific Responses Mediated by the Type IV Secretion System of Helicobacter pylori

doi: 10.1128/IAI.73.8.4643-4652.2005

Figure Lengend Snippet: Model of the roles of host cell factors required for the cellular responses mediated by the H. pylori TFSS. H. pylori translocates CagA, which is phosphorylated by Src family kinases (SFK). Tyrosine-phosphorylated CagA has been shown to interact with Csk and SHP-2, to inactivate Src family kinases, and to induce tyrosine dephosphorylation of cortactin and ezrin and rearrangements of the actin cytoskeleton. The TFSS also translocates peptidoglycan (PG), which activates the intracellular Nod1 receptor, activates the transcription factor NF-κB, and stimulates the expression and secretion of IL-8 and the upregulation of ICAM-1 on the cell surface. (A) Host cell factors appear to be required for CagA and peptidoglycan translocation. These factors could be inducers or cellular receptors of the TFSS. (B) Processing of CagA only occurs in some cell lines, indicating that specific cellular proteases are involved. (C) The hummingbird phenotype is only observed with AGS cells. Thus, some unknown host cell factors are apparently required for its induction. (D) HeLa and THP1 cells secrete some IL-8 in a TFSS-independent manner, indicating that another receptor can detect H. pylori in these cells.

Article Snippet: Cells were grown in the indicated media and subcultured every 2 to 3 days. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Cell line Reference or source a (no.) Cell type Growth medium b 293T ATCC (CRL 11268) Human embryonal kidney DMEM (Gibco BRL), 10% fetal bovine serum (FBS; Gibco BRL), 2 mM l -glutamine, 1 mM sodium pyruvate AGS ATCC (CRL 1739) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine CHO K1 ATCC (CCL 61) Chinese hamster ovary MEM alpha (Gibco BRL), 10% FBS Cos1 ATCC (CRL 1650) Green African monkey kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine GLC4 42 Human lung carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Hec1.b ATCC (HTP 113) Human endometrial adenocarcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine Hela ATCC (CCL 2) Human cervix epithelial carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HepG2 ATCC (HB 8065) Human hepatocellular carcinoma RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HL ECACC (96121720) Human lung, fibroblast-like RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine HT29 ATCC (HTB 38) Human colon adenocarcinoma Mc Coy's 5A (Gibco BRL), 10% FBS J774.A DSMZ (ACC 170) Mouse monocytic macrophages RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Kato3 ATCC (HTB 103) Human gastic carcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine L929 ATCC (CCL 1) Mouse fibroblasts RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MDCK ATCC (CCL 34) Canine kidney RPMI 1640 (Gibco BRL), 10% FBS, 2 mM l -glutamine MKN-28 27 Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine MKN-45 DSMZ (ACC 409) Human gastric adenocarcinoma RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine SR4987 DSMZ (ACC 323) Mouse stromal fibroblast-like Mc Coy's 5A (Gibco BRL), 10% FBS SYF+src ATCC (CRL 2498) Mouse fibroblasts overexpressing Src DMEM (Gibco BRL), 10% FBS, 2 mM l -glutamine, 1 mM sodium pyruvate THP1 ATCC (TIB 202) Human monocytes from acute monocytic leukemia RPMI 1640 (Gibco BRL), 10% heat-inactivated FBS, 2 mM l -glutamine Open in a separate window a ATCC, American Type Culture Collection ( www.atcc.org ); DSMZ, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH ( www.dsmz.de ); ECACC, European Collection of Cell Cultures ( www.ecacc.org.uk ). b DMEM, Dulbecco's modified Eagle's medium; MEM, minimum essential medium.

Techniques: De-Phosphorylation Assay, Expressing, Translocation Assay