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, Control, 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, Control, 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, Control, 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, Control, 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, Control, 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