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: