rabbit anti-egr-1 polyclonal antibody  (Santa Cruz Biotechnology)

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: Egr-1 protein was induced by HSV-1 infection 24 hours post treatment . a.) This figure is a western blot analysis showing the expression of Egr-1 protein (83 Kda) in infected VERO cells 24 hours post treatment. MOI of 1, 5, and 10 were used and more viruses led to more Egr-1 production. Noted that infection at higher moi (Lane 3) led to cell death therefore no significant increase of Egr-1 production was observed. b.) Egr-1 was induced by HSV-1 infection (MOI-5) in VERO cells and detectable levels were observed at 24, 48, and 72 hours post treatment. c.) Egr-1 induction was also seen in SIRC cells infected with a MOI of 5. d.) Uninfected HEK 293 cells express Egr1 (lane 1), and no induction was seen upon infection with HSV-1 MOI 10 (lane 2). Overexpression of Egr-1 was seen in HEK293 cells transfected with Egr1 expression vector (lane 3).

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Infection, Western Blot, Expressing, Over Expression, Transfection, Plasmid Preparation

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: Immunofluorescence microscopy indicates expression of Egr-1 in HSV-1 infected cells . Immunofluorescence data indicated that Egr-1(red) is expressed only in the cells infected with HSV-1 EGFP virus (green). Infection controls showed no EGFP signal or Egr-1, indicating the specificity of the antibody.

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Immunofluorescence, Microscopy, Expressing, Infection

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: Egr-1 mRNA was detected at 1 hour post treatment . VERO cells were infected with HSV-1 BE EGFP virus or with UV-inactivated virus at a MOI of 5. Total mRNA was isolated at 1, 2,3,12, and 24 hours post treatment and RT-PCR was performed.

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Infection, Isolation, Reverse Transcription Polymerase Chain Reaction

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: NFкB and CREB were recruited to the Egr-1 promoter upon infection . a.) Chip assay was performed using antibodies against NFкB and CREB. Lane1: Noninfected SIRC cells. Lane 2: SIRC cells infected with HSV-1 at a MOI of 5. Lane 3: HEK293 cells infected with virus at a MOI of 5. PCR was performed using primers for the Egr-1 promoter. For comparison, input controls (lanes 1-3) indicated the PCR products from samples prior to immunoprecipitation. b.) Over-expression of CREB protein (37 kda) was seen in VERO cells infected with HSV-1 (lane 2) when compared to the mock-infected control (lane 1).

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Infection, Immunoprecipitation, Over Expression

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: Dominant negative CREB inhibited the induction of Egr-1 upon HSV-1 infection . VERO cells were transfected with phMGFP alone and in combination with CREB dominant negative vectors pCREB133 or pKCREB. a.) The transfection efficiency of the three groups is similar as shown by the number of GFP-positive cells. b.) Cells infected with a control vector show induction of Egr-1 protein (red) whereas cells infected with the dominant negative mutants, CREB133(c) or KCREB (d) do not show the induction of the Egr-1 protein.

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Dominant Negative Mutation, Infection, Transfection, Plasmid Preparation

Journal: Virology Journal

Article Title: Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells

doi: 10.1186/1743-422X-8-262

Figure Lengend Snippet: NFкB inhibitors abolished the induction of Egr-1 upon infection . Egr-1 expression was not induced in cells pretreated with 1 μM of BAY 11-7082 (b) or 10 μM NBD binding peptide (c) after infection with HSV-1 compared the control (a)

Article Snippet: Rabbit anti-Egr-1 polyclonal antibody (Santa Cruz SC-110x) or cAMP response element binding (CREB) Ser-133 phosphorylation (Abcam#ab32096) was used at a dilution of 1:1,000.

Techniques: Infection, Expressing, Binding Assay

Journal: Molecular and cellular endocrinology

Article Title: DUAL INHIBITION OF ERK1/2 AND AKT PATHWAYS IS REQUIRED TO SUPPRESS THE GROWTH AND SURVIVAL OF ENDOMETRIOTIC CELLS AND LESIONS

doi: 10.1016/j.mce.2018.12.011

Figure Lengend Snippet: Details of the antibody used.

Article Snippet: Anti-human rabbit polyclonal EGR-1 , Cell Signaling , 4152 , 1:1000.

Techniques: Concentration Assay

Journal: Molecular cancer research : MCR

Article Title: A Key Role for Early Growth Response-1 and Nuclear Factor-κB in Mediating and Maintaining GRO/CXCR2 Proliferative Signaling in Esophageal Cancer

doi: 10.1158/1541-7786.MCR-08-0472

Figure Lengend Snippet: Expression of EGR-1 in normal and tumor esophageal tissues. A. Formalin-fixed, paraffin-embedded esophageal tumor tissues (top, II and IV) and adjacentnormal tissues (I and III) were subjected to immunohistochemical staining using polyclonal antibodies to human EGR-1 as described in Materials and Methods. EGR-1 was primarily located in the nucleus (see arrows in II and IV). Bar, 10 μm. b, basal cells of esophageal epithelium; m, mature epithelial cells; n, nucleus. An analysis of the stained sections shows that 80% (52 of 65) of the tumor tissues express EGR-1 at higher levels than observed in the corresponding normal tissues (right). B. Total RNA extracted from biopsies (normal and tumor tissue) obtained from patients diagnosed with esophageal squamous cell carcinoma was subjected to real-time RT-PCR, and the PCR products were analyzed by agarose gel electrophoresis; real-time RT-PCR was carried out with EGR-1-specific primers as described in Materials and Methods. Data represent the standardized expression of EGR-1 mRNA in esophageal squamous cell carcinoma tumor samples and adjacent uninvolved normal esophageal samples. Bars, SD for each sample analyzed in triplicate. *, P < 0.05, Student’s t test relative to normal control. Inset, real-time RT-PCR products separated on the agarose gel for EGR-1 and glyceraldehyde-3-phosphate dehydrogenase (lane 2) and the water blank control (lane 1).

Article Snippet: To detect the expression level of EGR-1 in positive clones selected with G418, cells were cultured on coverslips, fixed and permeabilized with methanol and 4% paraformaldehyde, and incubated with 1:100 dilution of rabbit anti-EGR-1 polyclonal antibody (Santa Cruz Biotechnology) in blocking buffer at 4°C overnight.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Immunohistochemical staining, Staining, Quantitative RT-PCR, Agarose Gel Electrophoresis

Journal: Molecular cancer research : MCR

Article Title: A Key Role for Early Growth Response-1 and Nuclear Factor-κB in Mediating and Maintaining GRO/CXCR2 Proliferative Signaling in Esophageal Cancer

doi: 10.1158/1541-7786.MCR-08-0472

Figure Lengend Snippet: Inhibitory effect of EGR-1 RNAi on cellular proliferation of esophageal cancer. A. To explore the potential role of EGR-1 in cellular proliferation of esophageal cancer, WHCO1 cells were stably transfected with either EGR-1 RNAi or EGR-1 sister control. Total cellular RNA was isolated from clones transfected with either EGR-1 RNAi or its sister control and subjected to real-time RT-PCR using primers to EGR-1. Data represent the standardized expression of EGR-1 mRNA in the indicated cell lines. Bars, SD for each sample analyzed in triplicate. *, P < 0.05, Student’s t test relative to normal control. B. Immunofluorescence staining was done using an EGR-1-specific antibody to determine the EGR-1 protein level and its subcellular localization as described in Materials and Methods. C. To detect the effect of silencing EGR-1 on the proliferation of WHCO1 cells, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was done as described in Materials and Methods and the absorbance was measured at 595 nm using a microtiter plate reader. Each point represents four analyses with the experiment repeated. D. To determine the effect of silencing EGR-1 on the progress of cell cycle, cell cycle analysis was carried out using propidium iodide staining as described in Materials and Methods and each sample was done in triplicate. *, P < 0.05.

Article Snippet: To detect the expression level of EGR-1 in positive clones selected with G418, cells were cultured on coverslips, fixed and permeabilized with methanol and 4% paraformaldehyde, and incubated with 1:100 dilution of rabbit anti-EGR-1 polyclonal antibody (Santa Cruz Biotechnology) in blocking buffer at 4°C overnight.

Techniques: Stable Transfection, Transfection, Isolation, Clone Assay, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Cell Cycle Assay

Journal: Molecular cancer research : MCR

Article Title: A Key Role for Early Growth Response-1 and Nuclear Factor-κB in Mediating and Maintaining GRO/CXCR2 Proliferative Signaling in Esophageal Cancer

doi: 10.1158/1541-7786.MCR-08-0472

Figure Lengend Snippet: Interference with EGR-1 signaling affects expression of key cell cycle regulators. Whole-cell lysates prepared from WHCO1 cells stably transfected with either EGR-1 RNAi, EGR-1 sister, GROα RNAi, GROβ RNAi, or vector control were separated on 7% to 15% gradient SDS-PAGE. Western blot analysis was carried out using antibodies specific to either EGR-1 or CDK4 or p27Kip1 as described in Materials and Methods.

Article Snippet: To detect the expression level of EGR-1 in positive clones selected with G418, cells were cultured on coverslips, fixed and permeabilized with methanol and 4% paraformaldehyde, and incubated with 1:100 dilution of rabbit anti-EGR-1 polyclonal antibody (Santa Cruz Biotechnology) in blocking buffer at 4°C overnight.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, SDS Page, Western Blot

Journal: Molecular cancer research : MCR

Article Title: A Key Role for Early Growth Response-1 and Nuclear Factor-κB in Mediating and Maintaining GRO/CXCR2 Proliferative Signaling in Esophageal Cancer

doi: 10.1158/1541-7786.MCR-08-0472

Figure Lengend Snippet: EGR-1 contributes to constitutive activation of NF-κB in esophageal cancer. A. Total RNA extracted from WHCO1 and WHCO1 clones containing GROα RNAi, GROβ RNAi, EGR-1 RNAi was subjected to real-time RT-PCR carried out with IκBα-specific primers as described in Materials and Methods, and the PCR products were analyzed by agarose gel electrophoresis (lane 1, water blank; lane 2, IκBα). Data represent the standardized expression of IκBα mRNA in WHCO1 transfected with vector and WHCO1 clones containing GROα RNAi, GROβ RNAi, and EGR-1 RNAi. Bars, SD for each sample measured in triplicate. *, P < 0.05, Student’s t test relative to vector control WHCO1. B. Total RNA extracted from EGR-1 stable clones and KYSE 450 transiently transfected with EGR-1 siRNA was subjected to real-time RT-PCR using p65-specific primers as described in Materials and Methods. Data represent the standardized expression of p65 mRNA in the indicated cell lines. Bars, SD for each sample measured in triplicate. C. Formalin-fixed, paraffin-embedded esophageal tumor tissues (II and IV) and adjacentnormal tissues (I and III) were subjected to immunohistochemical staining using monoclonal antibody to human p65 as described in Materials and Methods; p65 was located in both cytoplasm and nucleus. Bar, 10 μm. An analysis of the stained sections shows that 78% (48 of 61) of the tumor tissues express p65 at higher levels than observed in the corresponding normal tissues (bottom).

Article Snippet: To detect the expression level of EGR-1 in positive clones selected with G418, cells were cultured on coverslips, fixed and permeabilized with methanol and 4% paraformaldehyde, and incubated with 1:100 dilution of rabbit anti-EGR-1 polyclonal antibody (Santa Cruz Biotechnology) in blocking buffer at 4°C overnight.

Techniques: Activation Assay, Clone Assay, Quantitative RT-PCR, Agarose Gel Electrophoresis, Expressing, Transfection, Plasmid Preparation, Formalin-fixed Paraffin-Embedded, Immunohistochemical staining, Staining