Fig 2: Altered ARID2 expression modulates the growth of xenograft liver cancer tumor in mice. Male Balb/c nude mice were injected subcutaneously with 1×107 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.

Fig 3: 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.

Fig 4: 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.

Fig 5: MUC1-C induces PBRM1, ARID2, and BRD7 expression.A DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were analyzed for PBRM1, ARID2, and BRD7 mRNA levels by qRT-PCR using primers listed in Supplementary Table S1. The results (mean ± SD of 4 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). B Lysates from DU-145/tet-CshRNA and DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins. C Lysates from DU-145/CsgRNA, DU-145/MUC1sgRNA#1, and DU-145/MUC1sgRNA#2 cells were immunoblotted with antibodies against the indicated proteins. D LNCaP-AI/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were analyzed for PBRM1, ARID2, and BRD7 mRNA levels by qRT-PCR. The results (mean ± SD of 4 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). E LNCaP-AI/tet-CshRNA and LNCaP-AI/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins. F Lysates from LNCaP/tet-MUC1-C cells treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins. G Lysates from NCI-H660/CshRNA and NCI-H660/MUC1shRNA cells were immunoblotted with antibodies against the indicated proteins.