Journal: International Journal of Biological Sciences

Article Title: HIV-1-infected cell-derived exosomes promote the growth and progression of cervical cancer

doi: 10.7150/ijbs.38146

Figure Lengend Snippet: ARID2 is a direct downstream target of miR-155-5p in cervical cancer progression. (A) Target gene prediction for miR-1247-3p with two bioinformatics tools. (B) qRT-PCR and western blot assays of ARID2 expression in CaSki cells treated with miR-155-5p mimic or normal control. (C) The wild-type and a mutated type of the binding site between miR-155-5p and ARID2 3ʹ-UTR. (D) Relative luciferase activity of CaSki cells in the presence of the indicated treatments (E, F) Migration assay and qRT-PCR analysis of CaSki cells transfected with ARID2-specific siRNAs or control. (G) The effect of miR-155-5p on the migration ability of CaSki cells overexpressing ARID2.

Article Snippet: The primary antibodies and dilutions were as follows: anti-ARID2 (1:1000, Cell Signaling Technology, USA), anti-CD63, anti-CD9, anti-p-ERCC5, anti-CD81, anti-p-NF-KB, and anti-NF-KB (1:500, Proteintech, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Binding Assay, Luciferase, Activity Assay, Migration, Transfection

Journal: International Journal of Biological Sciences

Article Title: HIV-1-infected cell-derived exosomes promote the growth and progression of cervical cancer

doi: 10.7150/ijbs.38146

Figure Lengend Snippet: Exosomal miR-155-5p promotes cervical cancer progression via the ARID2-ERCC5-NF-κB signaling axis. (A) Volcano plot analysis of differentially expressed genes (fold change > 4; P < 0.05) after ARID2 knockdown. (B) Pathway network of differentially expressed genes. (C) Coexpression network of differentially expressed genes. (D) Immunoblotting assays of the indicated proteins in CaSki cells with the indicated treatments. (E, F) We used molecular docking to predict whether ARID2 and ERCC5 could bind. The predicted results show that ARID2 and ERCC5 bound by multiple potential interactions. Furthermore, we determined that ARID2 and ERCC5 bind to each other in cells by co-IP experiments.

Article Snippet: The primary antibodies and dilutions were as follows: anti-ARID2 (1:1000, Cell Signaling Technology, USA), anti-CD63, anti-CD9, anti-p-ERCC5, anti-CD81, anti-p-NF-KB, and anti-NF-KB (1:500, Proteintech, USA).

Techniques: Western Blot, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: A nuclear receptor facilitates differentiation of human PSCs into more mature hepatocytes

doi: 10.1101/2022.03.24.485641

Figure Lengend Snippet: Interactions between THRB and pBAF components in THRB-FLAG HepG2 cells. (A) Experimental design to identify THRB-binding proteins by immunoprecipitation with anti-FLAG antibody in THRB-FLAG HepG2 cells followed by mass-spectrometry and IP-Western blotting analysis. (B) Identification of high confidence nuclear proteins co-immunoprecipitated with THRB-FLAB ( P <0.01). (C) Control THRB-FLAG HepG2 cells or cells treated with 3 nM T3 for 1 day were lysed and subjected to anti-FLAG immunoprecipitation and Western blotting with pBAF components PBRM1 and ARID2.

Article Snippet: Western blotting was performed with a rabbit anti-PBRM1 antibody (Bethyl Laboratories A301-591A, 1:2000), a rabbit anti-ARID2 (Cell Signaling Technology 82342S 1:1000), or a mouse anti FLAG antibody (Sigma F1804 1:4000) based on previous methods ( ).

Techniques: Binding Assay, Immunoprecipitation, Mass Spectrometry, Western Blot

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Expression levels of mir-922 and ARID2 in patients with hepatocellular carcinoma.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: ARID2 is a potential target of miR-922 in liver cancer. Relative levels of FIGN and ARID2 expression levels in 10 pairs of liver cancer and adjacent non-tumor tissue were determined by reverse transcription-quantitative PCR and western blot analysis. WT or MT 3'UTR of FIGN and ARID2 were cloned into luciferase reporter vector to generate WT or MT FIGN and ARID2 luciferase reporter plasmids, respectively. Luciferase reporter gene assay was performed in MHCC97L cells following transfection with miR-922 mimics and plasmid. The presence of miR-922 in miRNA/AGO2 complex was determined by RNA immunoprecipitation using anti-AGO2 antibody. (A) FIGN and ARID2 mRNA transcripts in liver cancer tissue. FIGN and ARID2 protein expression levels in (B) liver cancer tissue and (C) HepG2, MHCC97H and MHCC97L and non-tumor hepatocyte THLE-2 cells. (D) Luciferase activity. (E) Representative images of agarose gel electrophoresis of PCR products. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. C, cancer tissue; P, para-carcinoma tissue; ARID2 , AT-rich interactive domain 2; miR, microRNA; FIGN, fidgetin, microtubule severing factor; WT, wild-type; MT, mutant; NC, negative control; ns, not significant.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Clone Assay, Luciferase, Plasmid Preparation, Reporter Gene Assay, Transfection, Immunoprecipitation, Activity Assay, Agarose Gel Electrophoresis, Mutagenesis, Negative Control

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression changes malignant behavior of liver cancer cells. The proliferation, clonogenicity, apoptosis, wound healing and invasion of HepG2 cells were determined following altered ARID2 expression. (A) Cell Counting Kit-8 assay determined cell proliferation. (Bi and Bii) Flow cytometry analysis of apoptotic cells. (Ci and Cii) Cell clonogenicity of HepG2 cells. (Di and Dii) Transwell assay analysis of HepG2 cell invasion. (Ei and Eii) Wound healing analysis of HepG2 cell proliferation and migration. Data are presented as the mean ± SD (n=3). *P<0.05, ***P<0.001. ARID2 , AT-rich interactive domain 2; OD, optical density; NC, negative control; sh, short hairpin; OE, overexpression.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing, Cell Counting, Flow Cytometry, Transwell Assay, Migration, Negative Control, Over Expression

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression modulates the growth of xenograft liver cancer tumor in mice. Male Balb/c nude mice were injected subcutaneously with 1×10 7 cells; 28 days later, tumor tissue was dissected and volume and weight were measured. The relative levels of miR-922 expression in tumor tissue were determined by reverse transcription-quantitative PCR and the levels of serum VEGF and TNF-α in mice were measured by ELISA. ARID2 , Bcl-2, Bax, PCNA, Cyclin D1, MMP3 and MMP9 expression levels in tumor tissue were characterized by immunohistochemistry. (A) Tumor-bearing mice. Tumor (B) volume and (C) weight. (D) Relative levels of miR-922 transcripts in tumor tissue. ELISA analysis of serum (E) VEGF and (F) TNF-α levels in mice. (G) Immunohistochemistry analysis of ARID2 , Bcl-2, Bax, PCNA, cyclin D1, MMP3 and MMP9 expression levels in tumor tissue. Data are presented as the mean ± SD (n=3). *P<0.05, ***P<0.001. ARID2, ARID2 , AT-rich interactive domain 2; PCNA, proliferating cell nuclear antigen; CON, control; NC, negative control; sh, short hairpin; OE, over-expression.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Negative Control, Over Expression

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression modulates miR-922 inhibitor-decreased malignant behavior of HepG2 cells. HepG2 cells were stably transfected with miR-922 inhibitor and transfected with plasmid for ARID2 expression or ARID2 -specific shRNA for ARID2 silencing. Control cells were transfected with vehicle. (A) Cell Counting Kit-8 determined cell proliferation. (Bi and Bii) Number of apoptotic HepG2 cells was analyzed by flow cytometry. (Ci and Cii) Clonogenicity of each group of HepG2 cells was analyzed by colony formation assay. (Di and Dii) Wound healing analysis of each group of HepG2 cells. (Ei and Eii) Transwell assay analysis determined the invasion of each group of HepG2 cells. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. ARID2 , AT-rich interactive domain 2; miR, microRNA; sh, short hairpin; OD, optical density; NC, negative control; OE, over-expression.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, shRNA, Cell Counting, Flow Cytometry, Colony Assay, Transwell Assay, Negative Control, Over Expression

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: ARID2 is a potential target of miR-922 in liver cancer. Relative levels of FIGN and ARID2 expression levels in 10 pairs of liver cancer and adjacent non-tumor tissue were determined by reverse transcription-quantitative PCR and western blot analysis. WT or MT 3'UTR of FIGN and ARID2 were cloned into luciferase reporter vector to generate WT or MT FIGN and ARID2 luciferase reporter plasmids, respectively. Luciferase reporter gene assay was performed in MHCC97L cells following transfection with miR-922 mimics and plasmid. The presence of miR-922 in miRNA/AGO2 complex was determined by RNA immunoprecipitation using anti-AGO2 antibody. (A) FIGN and ARID2 mRNA transcripts in liver cancer tissue. FIGN and ARID2 protein expression levels in (B) liver cancer tissue and (C) HepG2, MHCC97H and MHCC97L and non-tumor hepatocyte THLE-2 cells. (D) Luciferase activity. (E) Representative images of agarose gel electrophoresis of PCR products. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. C, cancer tissue; P, para-carcinoma tissue; ARID2 , AT-rich interactive domain 2; miR, microRNA; FIGN, fidgetin, microtubule severing factor; WT, wild-type; MT, mutant; NC, negative control; ns, not significant.

Article Snippet: The immunoblot membranes were blocked with 5% fat-free dry milk in TBST (0.1% Tween-20) buffer at 37°C for 2 h and incubated with primary antibodies overnight at 4°C ( ARID2 , 1:1,000, cat. no. #82342, Cell Signaling Technology, Inc.; FIGN, 1:1,000, cat. no. ab122238, Abcam).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Clone Assay, Luciferase, Plasmid Preparation, Reporter Gene Assay, Transfection, Immunoprecipitation, Activity Assay, Agarose Gel Electrophoresis, Mutagenesis, Negative Control

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Expression levels of mir-922 and ARID2 in patients with hepatocellular carcinoma.

Article Snippet: The supernatants were collected and incubated with ARID2 antibody (1:1,000, cat. no. #82342; Cell Signaling Technology, Inc.) and FIGN antibody (1:1,000, cat. no. ab122238, Abcam); overnight at 4°C.

Techniques: Expressing

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: ARID2 is a potential target of miR-922 in liver cancer. Relative levels of FIGN and ARID2 expression levels in 10 pairs of liver cancer and adjacent non-tumor tissue were determined by reverse transcription-quantitative PCR and western blot analysis. WT or MT 3'UTR of FIGN and ARID2 were cloned into luciferase reporter vector to generate WT or MT FIGN and ARID2 luciferase reporter plasmids, respectively. Luciferase reporter gene assay was performed in MHCC97L cells following transfection with miR-922 mimics and plasmid. The presence of miR-922 in miRNA/AGO2 complex was determined by RNA immunoprecipitation using anti-AGO2 antibody. (A) FIGN and ARID2 mRNA transcripts in liver cancer tissue. FIGN and ARID2 protein expression levels in (B) liver cancer tissue and (C) HepG2, MHCC97H and MHCC97L and non-tumor hepatocyte THLE-2 cells. (D) Luciferase activity. (E) Representative images of agarose gel electrophoresis of PCR products. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. C, cancer tissue; P, para-carcinoma tissue; ARID2 , AT-rich interactive domain 2; miR, microRNA; FIGN, fidgetin, microtubule severing factor; WT, wild-type; MT, mutant; NC, negative control; ns, not significant.

Article Snippet: The supernatants were collected and incubated with ARID2 antibody (1:1,000, cat. no. #82342; Cell Signaling Technology, Inc.) and FIGN antibody (1:1,000, cat. no. ab122238, Abcam); overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Clone Assay, Luciferase, Plasmid Preparation, Reporter Gene Assay, Transfection, Immunoprecipitation, Activity Assay, Agarose Gel Electrophoresis, Mutagenesis, Negative Control

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression changes malignant behavior of liver cancer cells. The proliferation, clonogenicity, apoptosis, wound healing and invasion of HepG2 cells were determined following altered ARID2 expression. (A) Cell Counting Kit-8 assay determined cell proliferation. (Bi and Bii) Flow cytometry analysis of apoptotic cells. (Ci and Cii) Cell clonogenicity of HepG2 cells. (Di and Dii) Transwell assay analysis of HepG2 cell invasion. (Ei and Eii) Wound healing analysis of HepG2 cell proliferation and migration. Data are presented as the mean ± SD (n=3). *P<0.05, ***P<0.001. ARID2 , AT-rich interactive domain 2; OD, optical density; NC, negative control; sh, short hairpin; OE, overexpression.

Article Snippet: The supernatants were collected and incubated with ARID2 antibody (1:1,000, cat. no. #82342; Cell Signaling Technology, Inc.) and FIGN antibody (1:1,000, cat. no. ab122238, Abcam); overnight at 4°C.

Techniques: Expressing, Cell Counting, Flow Cytometry, Transwell Assay, Migration, Negative Control, Over Expression

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression modulates the growth of xenograft liver cancer tumor in mice. Male Balb/c nude mice were injected subcutaneously with 1×10 7 cells; 28 days later, tumor tissue was dissected and volume and weight were measured. The relative levels of miR-922 expression in tumor tissue were determined by reverse transcription-quantitative PCR and the levels of serum VEGF and TNF-α in mice were measured by ELISA. ARID2 , Bcl-2, Bax, PCNA, Cyclin D1, MMP3 and MMP9 expression levels in tumor tissue were characterized by immunohistochemistry. (A) Tumor-bearing mice. Tumor (B) volume and (C) weight. (D) Relative levels of miR-922 transcripts in tumor tissue. ELISA analysis of serum (E) VEGF and (F) TNF-α levels in mice. (G) Immunohistochemistry analysis of ARID2 , Bcl-2, Bax, PCNA, cyclin D1, MMP3 and MMP9 expression levels in tumor tissue. Data are presented as the mean ± SD (n=3). *P<0.05, ***P<0.001. ARID2, ARID2 , AT-rich interactive domain 2; PCNA, proliferating cell nuclear antigen; CON, control; NC, negative control; sh, short hairpin; OE, over-expression.

Article Snippet: The supernatants were collected and incubated with ARID2 antibody (1:1,000, cat. no. #82342; Cell Signaling Technology, Inc.) and FIGN antibody (1:1,000, cat. no. ab122238, Abcam); overnight at 4°C.

Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Negative Control, Over Expression

Journal: Oncology Reports

Article Title: CREB1 acts via the miR-922/ ARID2 axis to enhance malignant behavior of liver cancer cells

doi: 10.3892/or.2021.8030

Figure Lengend Snippet: Altered ARID2 expression modulates miR-922 inhibitor-decreased malignant behavior of HepG2 cells. HepG2 cells were stably transfected with miR-922 inhibitor and transfected with plasmid for ARID2 expression or ARID2 -specific shRNA for ARID2 silencing. Control cells were transfected with vehicle. (A) Cell Counting Kit-8 determined cell proliferation. (Bi and Bii) Number of apoptotic HepG2 cells was analyzed by flow cytometry. (Ci and Cii) Clonogenicity of each group of HepG2 cells was analyzed by colony formation assay. (Di and Dii) Wound healing analysis of each group of HepG2 cells. (Ei and Eii) Transwell assay analysis determined the invasion of each group of HepG2 cells. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. ARID2 , AT-rich interactive domain 2; miR, microRNA; sh, short hairpin; OD, optical density; NC, negative control; OE, over-expression.

Article Snippet: The supernatants were collected and incubated with ARID2 antibody (1:1,000, cat. no. #82342; Cell Signaling Technology, Inc.) and FIGN antibody (1:1,000, cat. no. ab122238, Abcam); overnight at 4°C.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, shRNA, Cell Counting, Flow Cytometry, Colony Assay, Transwell Assay, Negative Control, Over Expression