mouse anti human atf3 monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Molecular Medicine Reports

Article Title: Overexpression of activating transcription factor 3 exerts suppressive effects in HepG2 cells

doi: 10.3892/mmr.2018.9707

Figure Lengend Snippet: Overexpression of ATF3 in HepG2 cells is determined by fluorescence microscope and western blot. A total of >90% cells with the green fluorescence were identified in the (A) OE and (B) NC groups. (C) No signal was measured in the control group, as determined by fluorescence microscope. Magnification, ×100. (D) ATF3 protein level in HepG2 cells was determined by western blot. It showed an increase in OE group compared with the NC and control groups was observed visually. ATF3, activating transcription factor 3; OE, overexpression; NC, negative control.

Article Snippet: Membranes were incubated with 5% non-fat milk for 1 h at room temperature and then with mouse monoclonal anti-human ATF3 antibody (1:100 dilution, ab58668; Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Over Expression, Fluorescence, Microscopy, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of activating transcription factor 3 exerts suppressive effects in HepG2 cells

doi: 10.3892/mmr.2018.9707

Figure Lengend Snippet: HepG2 proliferation ability following the overexpression of ATF3 is detected by MTT assay. (A) Growth curves of HepG2 cells in OE, NC and control groups. (B) Composite histograms of OD490/fold values in OE, NC and control groups. The OD490/fold value in OE group was significantly decreased compared with the NC and control groups from the third day following infection, while there was no significance between the NC and control groups (P=0.637). *P<0.05 vs. control and NC groups. Data are presented as the mean ± standard deviation. ATF3, activating transcription factor 3; OE, overexpression; NC, negative control; Con, control.

Article Snippet: Membranes were incubated with 5% non-fat milk for 1 h at room temperature and then with mouse monoclonal anti-human ATF3 antibody (1:100 dilution, ab58668; Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Over Expression, MTT Assay, Infection, Standard Deviation, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of activating transcription factor 3 exerts suppressive effects in HepG2 cells

doi: 10.3892/mmr.2018.9707

Figure Lengend Snippet: HepG2 transfer ability following overexpression of ATF3 is detected by Transwell assay. (A-C) Microscope images of Giesma-stained HepG2 transfer cells in (A) OE, (B) NC and (C) control groups. Magnification, ×100. (D) Histograms of OD570/OD490 values in OE, NC and control groups. Data are presented as the mean ± standard deviation. The transfer difference at the OE, NC and control groups was not marked, meanwhile no significance between groups was observed (P>0.05). ATF3, activating transcription factor 3; OE, overexpression; NC, negative control; Con, control; OD, optical density.

Article Snippet: Membranes were incubated with 5% non-fat milk for 1 h at room temperature and then with mouse monoclonal anti-human ATF3 antibody (1:100 dilution, ab58668; Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Over Expression, Transwell Assay, Microscopy, Staining, Standard Deviation, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of activating transcription factor 3 exerts suppressive effects in HepG2 cells

doi: 10.3892/mmr.2018.9707

Figure Lengend Snippet: HepG2 apoptotic rate following the overexpression of ATF3 was detected by Annexin V-allophycocyanin flow cytometry. (A-C) Peak pattern diagrams of the (A) OE, (B) NC and (C) control groups. The percentage of cells in the ‘M1’ region is representative of the apoptotic rate. The value in OE group was increased compared with the NC and control groups. (D) Peak pattern histograms of apoptotic rate in OE, NC and control groups. The HepG2 apoptotic rate in the OE group was significantly increased compared with the NC and control groups (*P<0.05), while no significance between the NC and control groups was observed (P=0.058). Data are presented as the mean ± standard deviation. ATF3, activating transcription factor 3; OE, overexpression; NC, negative control; Con, control.

Article Snippet: Membranes were incubated with 5% non-fat milk for 1 h at room temperature and then with mouse monoclonal anti-human ATF3 antibody (1:100 dilution, ab58668; Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Over Expression, Flow Cytometry, Standard Deviation, Negative Control

Journal: Molecular Medicine Reports

Article Title: Overexpression of activating transcription factor 3 exerts suppressive effects in HepG2 cells

doi: 10.3892/mmr.2018.9707

Figure Lengend Snippet: HepG2 cell cycle progression following overexpression of ATF3 was detected by propidium iodide staining. (A-C) Flow cytometry cycle diagram of (A) OE, (B) NC and (C) Con groups. DipG1 was representative of the proportion of cells in the G0/G1 phase; DipG2 was representative of the proportion of cells in the G2/M phase; and DipS was representative of the proportion of cells in the S phase. (D) Composite histograms of the cell proportion in G0/G1, S and G2/M phases among the OE, NC and Control groups. Data are presented as the mean ± standard deviation. The G0/G1 proportion in OE group was significantly increased compared with the NC and control groups, while the S and G2/M proportion were not significance compared with other groups. *P<0.05 vs. NC and control groups. (E) Histograms of the S+G2/M phase cell proportions in the OE, NC and control groups. Data are presented as the mean ± standard deviation. The proportion sum in OE group was significantly decreased compared with the NC and control groups, while no significance between the NC and control groups was observed. *P<0.05 vs. NC and control groups. ATF3, activating transcription factor 3; OE, overexpression; NC, negative control; Con, control.

Article Snippet: Membranes were incubated with 5% non-fat milk for 1 h at room temperature and then with mouse monoclonal anti-human ATF3 antibody (1:100 dilution, ab58668; Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Over Expression, Staining, Flow Cytometry, Standard Deviation, Negative Control

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: NDRG1 and ATF3 expression using Western blot and immunofluorescent staining in lung and lung cancer cell lines. A549 and H1299 cells were selected to investigate the function of NDRG1 and ATF3 in vitro. There was a medium level of NDRG1 and ATF3 expression in A549 cells but very weak expression in 1299 cells (1A). ATF3 was located in cytoplasm and nuclear in both A549 and 1299 cells, while NDRG1 was located in cytoplasm and nuclear in 1299 cells, and mainly in cytoplasm with weak expression in nuclear in A549 cells (1B).

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Expressing, Western Blot, Staining, In Vitro

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: Overexpression of NDRG1 and ATF3 in A549 cancer cells (A) and H1299 cells (B) after transfection of specific plasmids was confirmed by western blot.

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Over Expression, Transfection, Western Blot

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: Flow cytometry analysis of cisplatin-induced cytotoxicity in A549 cells. Overexpression of NDRG1 significantly reduced the cytotoxicity to A549 lung cancer cells induced by cisplatin, while overexpression of ATF3 significantly promoted cytotoxicity (P < 0.05).

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Flow Cytometry, Over Expression

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: Flow cytometry analysis of cisplatin-induced cytotoxicity in H1299 cells. Overexpression of ATF3 significantly promoted the cytotoxicity induced by cisplatin in H1299 cells ( p <0.05), while overexpression of NDRG1 didn't significantly impact on the cytotoxicity induced by cisplatin ( p >0.05).

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Flow Cytometry, Over Expression

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: Regulation of ATF3 and NDRG1 expression by NDRG1 overexpression. NDRG1 significantly downregulated ATF3 and P53 expression in A549 cells (* p < 0.05), but not in H1299 cells ( p >0.05).

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Expressing, Over Expression

Journal: International Journal of Medical Sciences

Article Title: NDRG1 Downregulates ATF3 and Inhibits Cisplatin-Induced Cytotoxicity in Lung Cancer A549 Cells

doi: 10.7150/ijms.28055

Figure Lengend Snippet: Regulation of P53 expression by ATF3 overexpression. ATF3 significantly upregulated P53 expression (* p < 0.05) but did not affect NDRG1 expression in A549 cells ( p > 0.05). ATF3 did not regulate P53 and NDRG1 expression in H1299 cells ( p > 0.05).

Article Snippet: Cells were fixed with 4% paraformaldehyde and then blocked with 1% BSA, followed by incubation with rabbit anti-human NDRG1 polyclonal antibody (ab63989, Abcam, HK; dilution 1:100) and mouse anti-human ATF3 monoclonal antibody (sc-81189, Santa Cruz, USA; dilution 1:100) overnight at 4°C.

Techniques: Expressing, Over Expression

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) Expression of ATF3 in the progression from normal epithelium to carcinoma of esophagus. Scale bar, 50μm. (B) Expression of ATF3 protein in four randomly selected, paired ESCC samples and matched normal tissues was analyzed by Western blotting. Signal intensity for the expression of ATF3 was quantified by densitometric scanning and normalized by internal control (β-actin). (C) ATF3 levels in whole-cell extracts were determined in various ESCC cell lines and immortalized esophageal epithelial cell lines. EC171, EC9706, KYSE150, EC109 and KYSE510 were ESCC cell lines. NE1, NE2 and NEcA6 were immortalized esophageal epithelial cell lines. (D) Immunofluorescence analysis of ATF3 expression in KYSE150 cells, an ESCC cell lines with high-expression of ATF3 (400×). (E) Comparison for the invasive capability of cells lines with different ATF3 expression level.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) Forced expression of ATF3 in EC109 and KYSE510 ESCC cell lines were addressed by Western blotting analysis. ATF3-1# and ATF3-2# were two different ATF3-transfected cell clones; Vector was cells transfected with vector control. (B) Colony formation assay was used to evaluate the growth of ATF3-expressing cells. (C) Invasiveness assay was used to determine the effect of ATF3 forced expression on cell invasion. Representative tumor cells invaded were photographed (400×), data represent mean ± SD of triplicates. (D) RNAi-mediated knockdown (siATF3) and re-expression of ATF3 (siATF3/ATF3) in KYSE150 cells were determined by Western blotting. Colony formation assay (E) and invasiveness assay (F) were employed to address the alterations of cell growth and invasion upon ATF3 knockdown and re-expression.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: Expressing, Western Blot, Transfection, Clone Assay, Plasmid Preparation, Colony Assay

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) & (B) ATF3 forced expression EC109 cells and the control cells were implanted subcutaneously in nude mice. Tumor volume in different time points (A) and average weight of the tumors (B) were analyzed. (C) & (D) & (E) ATF3 forced expression EC109 cells and the control cells were inoculated via tail veins of the SCID mice. (C) The photomicrographs of H&E-stained lung tissues. Representative fields were shown and metastatic colonizations were marked with arrows. (D) and (E) Quantitative analysis of the number of surface lung metastasis colonization. *, P < 0.05.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: Expressing, Staining

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) Western blotting analysis of MMP-2 expression in ATF3 forced expression or knockdown cells. β-actin served as a loading control. (B) Zymography of the conditioned medium for the activity of MMP-2. The coomassie staining of total protein in conditioned media were used to demonstrate that equal numbers of cells were present during the conditioning of the media. (C) Immunohistochemical staining of MMP-2 in the subcutaneous tumor tissues. Scale bar, 50μm. (D) Transcriptional level of MMP-2 was addressed by real time RT-PCR. (E) SiRNA targeted ATF3 (siATF3) and siRNA targeted MMP-2 (siMMP-2) were co-transfected into KYSE150 cells. (F) MMP-2 silencing inversed the increased cell invasion mediated by ATF3 knockdown.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: Western Blot, Expressing, Zymography, Activity Assay, Staining, Immunohistochemical staining, Quantitative RT-PCR, Transfection

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) Transfected EC109 cells and KYSE510 cells were treated with MG132 or NH 4 Cl, and then harvested for Western blotting analysis of MMP-2. (B) Expressions of MDM2, total P53 and nuclear P53 in ATF3 forced expression EC109 cells was addressed by Western blotting. β-actin and Nucleoporin p62 served as loading controls. (C) Increased expression of MDM2 was confirmed in the subcutaneous tumor tissues by immunohistochemical staining. Scale bar, 50μm. (D) ATF3 forced expression EC109 cells and the control cells were treated with MG132, MDM2 inhibitor or both MDM2 inhibitor and MG132 and then used for co-immunoprecipitation. The ubiquitinated and the total level of MMP-2 were addressed by Western blotting. (E) The reciprocal protein complexes involving ATF3, MDM2 and MMP-2 in the ATF3-expressing cells. An antibody against MMP-2 was subjected to co-IP experiment and expression of ATF3, MDM2 or MMP-2 was addressed. β-actin was served as a loading control.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: Transfection, Western Blot, Expressing, Immunohistochemical staining, Staining, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) Cytotoxicity of Cisplatin treatment on EC109 cells was determined by MTT assay. (B) Time course analysis for the expressions of ATF3, MDM2, MMP-2, P53 and p-ERK1/2 in EC109 cells treated with Cisplatin (4μg/ml). (C) Effect of Cisplatin on the formation of the ATF3/MDM2/MMP-2 complex. (D) ATF3 was silenced by RNAi method in the Cisplatin-treated cells. (E) MTT assay was employed to determine the effect of ATF3 silencing on Cisplatin-induced inhibition of cell growth. (F) Role of ATF3 in the Cisplatin-induced inhibition of cell invasion was addressed by invasiveness assay.

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: MTT Assay, Inhibition

Journal: Oncotarget

Article Title: ATF3 functions as a novel tumor suppressor with prognostic significance in esophageal squamous cell carcinoma

doi:

Figure Lengend Snippet: Proposed model illustrating opposing regulatory influences of ATF3 on MMP-2 degradation and cancer cell invasion and metastasis

Article Snippet: Rabbit anti-human ATF3 polyclonal antibody (Rockland, Pennsylvania, USA) and Mouse anti-human ATF3 polyclonal antibody (ABGENT, San Diego, USA) were used.

Techniques: