Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 2. SiRNA‑ATF2 transfection facilitated the anti‑proliferation effect of sorafenib. Data are presented as the mean ± standard error of the mean of 6 repeats. *P<0.05 vs. control, #P<0.05 vs. vector+sorafenib. ATF2, activating transcription factor‑2; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Transfection, Control, Plasmid Preparation, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 1. SiRNA‑ATF2 effectively reduced ATF2 expression. (A) Representative images of phase‑contrast and (B) fluorescence microscopy following siRNA‑ATF2 transfection and (C) ATF2 mRNA expression following transfection. Data are presented as the mean ± standard error of the mean of 6 repeats. *P<0.05 vs. control. ATF2, activating transcription factor‑2; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Expressing, Fluorescence, Microscopy, Transfection, Control, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 3. SiRNA‑ATF2 facilitated the apoptosis induced by sorafenib. (A) Flow cytometry was used to determine apoptosis. (B) Quantification of flow cytometry. Data are presented as the mean ± standard error of the mean of 6 repeats. *P<0.05 vs. control, #P<0.05 vs. vector+sorafenib. ATF2, activating transcription factor‑2; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Flow Cytometry, Control, Plasmid Preparation, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 4. SiRNA‑ATF2 facilitated the anti‑migration effect of sorafenib treatment. ATF2, activating transcription factor‑2; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Small Interfering RNA

Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 5. SiRNA‑ATF2 facilitated the anti‑invasion effect of sorafenib. (A) Representative images of the invaded cells. (B) Quantification of invasive cell number. Data are presented as the mean ± standard error of the mean of 6 repeats. *P<0.05 vs. control; #P<0.05 vs. vector+sorafenib. ATF2, activating transcription factor‑2; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Control, Plasmid Preparation, Small Interfering RNA

Journal: Molecular medicine reports

Article Title: Silencing activating transcription factor 2 promotes the anticancer activity of sorafenib in hepatocellular carcinoma cells.

doi: 10.3892/mmr.2018.8921

Figure Lengend Snippet: Figure 6. Sorafenib and/or siRNA‑ATF2 reduce ATF2 and JNK3 expression and enhance TNF‑α expression. mRNA expression of (A) ATF2, JNK3 and TNF‑α. (B) Representative western blot images for ATF2, p‑ATF2, TNF‑α, JNK3 and p‑JNK3. Quantification of (C) ATF2 and p‑ATF2, (D) TNF‑α and (E) p‑JNK3/JNK3 protein expression levels. Data are presented as the mean ± standard error of the mean of 6 repeats. *P<0.05 vs. control, #P<0.05 vs. vector+sorafenib. p, phosphory lated; ATF2, activating transcription factor‑2; siRNA, small interfering RNA; TNF‑α, tumor necrosis factor‑α; JNK3, c‑Jun N‑terminal kinase.

Article Snippet: The membranes were blocked in 5% skim milk for 2 h at room temperature and incubated with the following primary antibodies overnight at 4 ̊C: JNK3 (cat. no. ab76572; 1:1,000; Abcam, Cambridge, UK), ATF2 (cat. no. bs-0518R; 1:400; BIOSS, Beijing, China), GAPDH (cat. no. A007; 1:1,000; ABclonal Biotech Co., Ltd., Woburn, MA, USA), TNF-α (cat. no. BA14901; 1:800; BIOSS), phosphorylated (p)-JNK3 (cat. no. ab76572; 1:1,000; Abcam), p‐ATF2 (cat. no. BS‐84449R; 1:400, BIOSS) overnight at 4 ̊C.

Techniques: Expressing, Western Blot, Control, Plasmid Preparation, Small Interfering RNA

Journal: Communications biology

Article Title: The ATF2/miR-3913-5p/CREB5 axis is involved in the cell proliferation and metastasis of colorectal cancer.

doi: 10.1038/s42003-023-05405-w

Figure Lengend Snippet: Fig. 5 ATF2 transcriptionally regulates miR-3913-5p by binding to its promoter. a The CREB5 interacted with transcription factor proteins. b QPCR analyses of miR-3913-5p expression in CRC cells following treatment of ATF2, ATF7 or BATF3 interference. Student’s t test; *p > 0.05; ***p < 0.01; ****p < 0.001. c Cell lysates were immunoprecipitated by anti-CREB5 antibody or anti-ATF2 antibody. IgG was used as a control. Western blot analyses were performed using anti- CREB5 antibody or anti-ATF2 antibody. d The transcriptional factor ATF2-binding motif was predicted by informatics analysis. e Schematic illustration of the miR-3913-5p promoter with a potential ATF2 binding site. f Amplification of fragments containing ATF2-binding sites after ChIP assays using anti-ATF2 antibody is indicated in PCR gel. g Luciferase reporter assays were used to confirm the ATF2 binding to the promoter of miR- 3913-5p. Student’s t test; **p < 0.05; ***p < 0.01; ****p < 0.001.

Article Snippet: DNA fragments ranging from 400 to 500 bp were yielded via sonication and subjected to the immunoprecipitation process with anti-ATF2 antibody (Cell Signaling, #35031).

Techniques: Binding Assay, Expressing, Immunoprecipitation, Control, Western Blot, Luciferase

Journal: Communications biology

Article Title: The ATF2/miR-3913-5p/CREB5 axis is involved in the cell proliferation and metastasis of colorectal cancer.

doi: 10.1038/s42003-023-05405-w

Figure Lengend Snippet: Fig. 6 Inhibiting miR-3913-5p reverses the suppressions of CRC cell proliferation, migration and invasion induced by ATF2 interference. Colony formation assays (a) and EdU assays (b) revealed that inhibition of miR-3913-5p weakened the suppression of CRC cells growth by ATF2 interference. Student’s t test; ***p < 0.01; ****p < 0.001. Transwell assays showed that the suppression of CRC cells migration (c) and invasion (d) abilities induced by ATF2 interference was reversed by miR-3913-5p inhibitor. Student’s t test; ***p < 0.01; ****p < 0.001. Scale bars, 50 μm in (b–d).

Article Snippet: DNA fragments ranging from 400 to 500 bp were yielded via sonication and subjected to the immunoprecipitation process with anti-ATF2 antibody (Cell Signaling, #35031).

Techniques: Migration, Inhibition

Journal: Communications biology

Article Title: The ATF2/miR-3913-5p/CREB5 axis is involved in the cell proliferation and metastasis of colorectal cancer.

doi: 10.1038/s42003-023-05405-w

Figure Lengend Snippet: Fig. 7 miR-3913-5p is negatively correlated with ATF2 and CREB5 expression. Western blot assays (a) and qPCR assays (b) showed the CREB5, ATF2 and miR-3913-5p expression in 12 paired CRC tissues and their matched normal tissues. Student’s t test; **p < 0.05; ***p < 0.01; ****p < 0.001. c QPCR analyses of the CREB5 and ATF2 expression in 91 paired CRC tissues and their adjacent normal tissues. d The CREB5 and ATF2 expression levels in 91 pairs of CRC specimens and normal tissues. Student’s t test; **p < 0.05; ***p < 0.01. The expression correlations between CREB5 and ATF2 (e), between miR- 3913-5p and CREB5 (f), between miR-3913-5p and ATF2 (g) in CRC tissues. h ISH analyses of miR-3913-5p and IHC analyses of CREB5 and ATF2 in the CRC tissues and the corresponding normal tissues. i The illustration depicting the mechanism of ATF2/miR-3913-5p/CREB5 axis in CRC. Scale bars, 100 μm in (h).

Article Snippet: DNA fragments ranging from 400 to 500 bp were yielded via sonication and subjected to the immunoprecipitation process with anti-ATF2 antibody (Cell Signaling, #35031).

Techniques: Expressing, Western Blot