Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Expression of dbpA in colorectal tumor and adjacent normal tissue samples.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Expression of DNA binding protein A (dbpA) in tissues obtained from patients with colorectal cancer (CRC) and various CRC cell lines using immunohistochemistry (IHC). T, tumor tissues; A, adjacent normal tissues; C, cytoplasm; and T1-T4, invasion depth. (A) dbpA expression in tumors and adjacent normal tissues (×100 magnification). (B) The levels of dbpA expression in different invasion depths (×100 magnification). RT-qPCR analysis of the mRNA expression of the dbpA in various human CRC cell lines. (C) Western blot analysis of dbpA protein levels. GAPDH was used as an internal control. Data are shown as the means ± standard deviation (SD).

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Expressing, Binding Assay, Immunohistochemistry, Quantitative RT-PCR, Western Blot, Control, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Association between dbpA expression and clinicopathologic factors in patient with CRC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Expressing, Significance Assay

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: DNA binding protein A (dbpA) knockdown by lentiviral-mediated RNAi. (A) Representative images of SW620 cells transfected with shRNA-dbpA-Lv, shNC-dbpA-Lv, or untransfected cells (CON) for 72 h (×100 magnification). GFP expression indicated that the cells had been successfully infected with the shRNA. (B) RT-qPCR analysis of dbpA mRNA levels in cells treated as in (A). (C) Western blot analysis showing dbpA protein levels in cells treated as in (A). GAPDH was used as an internal control. Data are shown as the means ± standard deviation (SD); ** p<0.01 vs. NC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Binding Assay, Knockdown, Transfection, shRNA, Expressing, Infection, Quantitative RT-PCR, Western Blot, Control, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Effect of DNA binding protein A (dbpA) silencing on SW620 cell proliferation. (A) MTT assay measuring cell proliferation of lentivirus-transfected SW620 cells. (B) Quantification of clone number in SW620 cells transfected with shRNA-dbpA-Lv, shNC-dbpA-Lv, or empty vector-transfected cells (CON). (C) Representative images of colony formation assay for SW620 cells in a 6-well plate. (D) Representative images per colony for SW620 cells under a bright microscope. Data are shown as the means ± standard deviation (SD); * p<0.05, ** p<0.01 vs. NC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Binding Assay, MTT Assay, Transfection, shRNA, Plasmid Preparation, Colony Assay, Microscopy, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Effect of DNA binding protein A (dbpA) silencing on cell cycle progression in SW620 cells. (A) Representative images of cell cycle analysis at 72 h by flow cytometry. (B) Cell cycle distribution in SW620 cells transfected with shRNA-dbpA-Lv, shNC-dbpA-Lv, or empty vector. Data are shown as the means ± standard deviation (SD); ** p<0.01 vs. NC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Binding Assay, Cell Cycle Assay, Flow Cytometry, Transfection, shRNA, Plasmid Preparation, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Apoptosis induction in SW620 cells transfected with short hairpin RNA (shRNA)-DNA binding protein A (dbpA)-Lv, shNC-dbpA-Lv, or empty vector. (A) Cell apoptosis was analyzed by flow cytometry. (B) Cell apoptosis in SW620 cells treated as in (A). Data are shown as the means ± standard deviation (SD); ** p<0.01 vs. NC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Transfection, shRNA, Binding Assay, Plasmid Preparation, Flow Cytometry, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: RNAi-mediated downregulation of DNA binding protein A inhibits tumorigenesis in colorectal cancer

doi: 10.3892/ijmm.2016.2662

Figure Lengend Snippet: Effect of DNA binding protein A (dbpA) knockdown on tumorigenesis in nude mice. (A) Curve of tumor volume was assessed by caliper measurements. (B) Mass weight of tumor at the 35th day after inoculation. (C) Representative images of mice and tumors from each group. (D) dbpA expression was detected by western blot analysis from isolated tumors. GAPDH was used as an internal control. Data are shown as the means ± standard deviation (SD); * p<0.05, ** p<0.01 vs. NC.

Article Snippet: After blocking with 5% milk, the filter was incubated overnight with a primary rabbit polyclonal anti-dbpA antibody (ab48952; Boster Biological Technology, Ltd.) at 1:500 dilutions for 1 h. The samples were then incubated with the HRP-conjugated secondary antibodies (ab6721) at 1:1,000, and the bands were detected by enhanced chemiluminescence (both from Amersham Biosciences, Piscataway, NJ, USA), and then analyzed using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Binding Assay, Knockdown, Expressing, Western Blot, Isolation, Control, Standard Deviation