Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Bacterial strains used in this study .

Article Snippet: The human pharyngeal epithelial cell line FaDu (ATCC HTB-43), the human colon epithelial cell line Caco-2 (ATCC HTB-37) and the human cervical epithelial cell line ME180 (ATCC HTB-33) were cultured in DMEM + GlutaMAX (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (Sigma Aldrich).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Cell type specificity in the bacteria-mediated induction of EGR1. (A–E) Cell line specific induction of EGR1 was investigated during bacterial infection of epithelial cell lines of a pharyngeal (FaDu), gastric (AGS), intestinal (Caco-2), and cervical origin (ME180). Expression of EGR1 was monitored using qPCR at 1, 2, 4, and 6 h post infection. (F–J) Bacterial attachment levels to epithelial cells at different time points. Bacteria were diluted and plated for viable counts to determine CFU/ml. Bacteria were added at a MOI of 100 for all experiments. Cells were infected with N. meningitidis (Nm-C), S. pyogenes (Sp-M6), H. pylori (Hp-J99), N. gonorrhoeae (Ng) or S. Enteritidis (SE-3934).

Article Snippet: The human pharyngeal epithelial cell line FaDu (ATCC HTB-43), the human colon epithelial cell line Caco-2 (ATCC HTB-37) and the human cervical epithelial cell line ME180 (ATCC HTB-33) were cultured in DMEM + GlutaMAX (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (Sigma Aldrich).

Techniques: Bacteria, Infection, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Bacterial viability and direct contact between bacteria and the host cells affect upregulation of EGR1 . Induction of EGR1 was monitored by qPCR after infection of the host epithelial cells with live or dead bacteria. Dead bacteria were obtained by heat treatment at 95°C for 10 min. Viable counts were used to ensure complete killing of the bacteria. The role of bacterial contact with the epithelial cells in the EGR1 induction was studied using a 0.4 μm Millicell filter, which helps to physically separate the bacteria and host cells, but still allows diffusion of secreted factors in the cell growth media (Sup.). (A) Pharyngeal FaDu cells infected with N. meningitidis (Nm-C). (B) Pharyngeal FaDu cells infected with S. pyogenes (Sp-M6). (C) Gastric AGS cells infected with H. pylori (Hp-J99). (D) Cervical ME180 cells infected with N. gonorrhoeae (Ng). (E) Intestinal Caco-2 cells infected with S. Enteritidis (SE3439). Bacteria were added to a MOI of 100 in all experiments. The data was analyzed at 2 h post infection for Nm-C, Sp-M6, SE3439, and Hp-J99. The data was analyzed at 4 h post infection of Ng.

Article Snippet: The human pharyngeal epithelial cell line FaDu (ATCC HTB-43), the human colon epithelial cell line Caco-2 (ATCC HTB-37) and the human cervical epithelial cell line ME180 (ATCC HTB-33) were cultured in DMEM + GlutaMAX (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (Sigma Aldrich).

Techniques: Bacteria, Infection, Diffusion-based Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: EGR1 is primarily activated by the EGFR–ERK1/2 pathway upon bacterial infection . Host epithelial cells were pretreated with PD153035, PD184352, SP600125, P38-IV and PKA-14:22 (inhibiting EGFR, ERK1/2, JNK, p38 and PKA, respectively) 1 h prior to infection. Bacterial infection of the host epithelial cells was carried out by co-incubation with the inhibitors for 2 h, except infection with N. gonorrhoeae that continued for 4 h. The expression of EGR1 was analyzed by qPCR. Expression of cells treated with DMSO was set to 1. (A) FaDu infected with N. meningitidis serogroup C (Nm-C). (B) FaDu infected with S. pyogenes serogroup M6 (Sp-M6). (C) AGS infected with H. pylori J99 (Hp-J99). (D) ME180 infected with N. gonorrhoea MS11 (Ng). (E) Caco-2 infected with S. Enteritidis (SE-3934). Bacteria were added to a MOI of 100 in all experiments. The white bars represent uninfected controls. The colored bars represent the infected samples. The significant difference between the infected control (DMSO) and the infected samples is marked with asterisk.

Article Snippet: The human pharyngeal epithelial cell line FaDu (ATCC HTB-43), the human colon epithelial cell line Caco-2 (ATCC HTB-37) and the human cervical epithelial cell line ME180 (ATCC HTB-33) were cultured in DMEM + GlutaMAX (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (Sigma Aldrich).

Techniques: Infection, Incubation, Expressing, Bacteria, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Integrin mediated signaling in bacteria induced EGR1 expression . The host epithelial cells were transfected with control siRNA (si-NT), siRNA targeted against β1-integrin (si-Integrin) or directed against integrin-linked kinase (si-ILK) for 60–68 h. The cells were then infected with bacteria with a MOI of 100 for 2 h, except the infection with N. gonorrhoeae that continued for 4 h. EGR1 expression was analyzed by qPCR. The graphs represent fold difference in EGR1 expression between infected and uninfected epithelial cells. (A) FaDu infected with N. meningitidis serogroup C (Nm-C). (B) FaDu infected with S. pyogenes serogroup M6 (Sp-M6). (C) MKN45 infected with H. pylori J99 (Hp-J99). (D) ME180 infected with N. gonorrhoea MS11 (Ng). (E) Caco-2 infected with S. Enteritidis (SE-3934).

Article Snippet: The human pharyngeal epithelial cell line FaDu (ATCC HTB-43), the human colon epithelial cell line Caco-2 (ATCC HTB-37) and the human cervical epithelial cell line ME180 (ATCC HTB-33) were cultured in DMEM + GlutaMAX (Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (Sigma Aldrich).

Techniques: Bacteria, Expressing, Transfection, Control, Infection

Journal: Carcinogenesis

Article Title: Stimulation of CEACAM1 expression by 12-O-tetradecanoylphorbol-13-acetate (TPA) and calcium ionophore A23187 in endometrial carcinoma cells.

doi: 10.1093/carcin/bgi275

Figure Lengend Snippet: Fig. 6. Luciferase (Luc) activity (arbitrary units) of the CEACAM1 deletion mutants P1–P4 and the vector control in TPA/calcium ionophore stimulated and unstimulated Hec1B (H) and Skut1B (S) cells (A) and stimulated and unstimulated SW480 (SW) and T47D (T) cells (B), respectively. Putative binding sites of the transcription factors AP-1 (D), AP-2 (*), AP-3 (^) and NFkB (*) and binding sites of already identified transactivating factors are indicated (33,51).

Article Snippet: Hec1B human endometrial adenocarcinoma cells, endometrial mixed mesenchymal Skut1B cells, SW480 colon carcinoma cells, T47D breast carcinoma cells and ECV304 bladder carcinoma cells were obtained from ATCC (Rockville, MD).

Techniques: Luciferase, Activity Assay, Plasmid Preparation, Control, Binding Assay

Journal: BioMed Research International

Article Title: A pH-Sensitive, Biobased Calcium Carbonate Aragonite Nanocrystal as a Novel Anticancer Delivery System

doi: 10.1155/2013/587451

Figure Lengend Snippet: In vitro cytotoxicity study of MDA MB231 cells after 24, 48, and 72 hours of exposure to free DOX and the CaCO 3 /DOX nanocrystals.

Article Snippet: Breast cancer cells, MDA MB231, were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a DMEM medium supplemented with 10% fetal bovine serum, L-glutamine (15 mmol/L), penicillin (100 U/mL), and streptomycin (100 μ g/mL).

Techniques: In Vitro

Journal: BioMed Research International

Article Title: A pH-Sensitive, Biobased Calcium Carbonate Aragonite Nanocrystal as a Novel Anticancer Delivery System

doi: 10.1155/2013/587451

Figure Lengend Snippet: Modified neutral red/trypan blue exclusion assay. MDA MB231 cells after 24, 48, and 72 h of exposure to free DOX and CaCO 3 /DOX nanocrystals.

Article Snippet: Breast cancer cells, MDA MB231, were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a DMEM medium supplemented with 10% fetal bovine serum, L-glutamine (15 mmol/L), penicillin (100 U/mL), and streptomycin (100 μ g/mL).

Techniques: Modification, Trypan Blue Exclusion Assay

Journal: BioMed Research International

Article Title: A pH-Sensitive, Biobased Calcium Carbonate Aragonite Nanocrystal as a Novel Anticancer Delivery System

doi: 10.1155/2013/587451

Figure Lengend Snippet: LDH release of MDA MB231 cells after 24, 48, and 72 hours of exposure to free DOX and the CaCO 3 /DOX nanocrystals.

Article Snippet: Breast cancer cells, MDA MB231, were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a DMEM medium supplemented with 10% fetal bovine serum, L-glutamine (15 mmol/L), penicillin (100 U/mL), and streptomycin (100 μ g/mL).

Techniques:

Journal: BioMed Research International

Article Title: A pH-Sensitive, Biobased Calcium Carbonate Aragonite Nanocrystal as a Novel Anticancer Delivery System

doi: 10.1155/2013/587451

Figure Lengend Snippet: Images of MDA MB231 cells treated with different agents at 24, 48, and 72 h of incubation.

Article Snippet: Breast cancer cells, MDA MB231, were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a DMEM medium supplemented with 10% fetal bovine serum, L-glutamine (15 mmol/L), penicillin (100 U/mL), and streptomycin (100 μ g/mL).

Techniques: Incubation

Journal: BioMed Research International

Article Title: A pH-Sensitive, Biobased Calcium Carbonate Aragonite Nanocrystal as a Novel Anticancer Delivery System

doi: 10.1155/2013/587451

Figure Lengend Snippet: Fluorescent images of MDA MB231 cells stained with Acridine orange (AO): (a) control, (b) treated with free DOX, and (c) CaCO 3 /DOX nanocrystals.

Article Snippet: Breast cancer cells, MDA MB231, were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a DMEM medium supplemented with 10% fetal bovine serum, L-glutamine (15 mmol/L), penicillin (100 U/mL), and streptomycin (100 μ g/mL).

Techniques: Staining, Control

Journal: Cancer

Article Title: Adenosine diphosphate-ribosylation factor 6 is required for epidermal growth factor-induced glioblastoma cell proliferation.

doi: 10.1002/cncr.24550

Figure Lengend Snippet: FIGURE 1. The expression of epidermal growth factor receptor (EGFR) and adenosine diphosphate-ribosylation factor 6 (ARF6) in human glioma tissue samples and cell lines is illustrated. Representative results are shown from (A) 16 high-grade glioma tis- sues (reverse transcriptase-polymerase chain reaction [RT-PCR]); (B) 6 low-grade glioma tissues (RT-PCR); and (C) 6 glioblas- toma cell lines, including U87, U138, U373, LN308, SW1088, and SW1783 (Western blot analysis). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and glial fibrillary acidic protein (GFAP) were used simultaneously as internal controls for equal loading of PCR products. N1 and N2 denote 2 samples of normal brain tissue. The relative band intensities were measured using densi- tometry and were normalized against their respective GAPDH signals. Data shown are the mean standard deviation of the in- tensity levels from 3 independent PCR amplifications. An asterisk indicates P < .05 for ARF6 in glioma tissues versus ARF6 in normal brain tissues; pound sign, P < .05 for EGFR in glioma tissues versus EGFR in normal brain tissues.

Article Snippet: U87, U138, U373, SW1088, and SW1783 cells were obtained from the American Type Culture Collection (ATCC) (Manassas, Va), and LN308 cells were a gift from Dr. M. E. Hegi (University Hospital Lausanne, Lausanne, Switzerland).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Standard Deviation