Journal: International Journal of Clinical and Experimental Pathology

Article Title: CBX8, a novel DNA repair protein, promotes tumorigenesis in human esophageal carcinoma

doi:

Figure Lengend Snippet: Expression of CBX8 in esophageal tissues and cell lines. A. CBX8 mRNA levels were measured in 59 pairs of esophageal carcinoma and corresponding adjacent normal tissues by real-time RT-PCR, where 18S was used as an internal reference. The line in each box is the median -ΔCt value, while the upper and lower edges of each box show the 75th and 25th percentile, respectively. P value was calculated by paired t-test (P<0.01). B. The difference in CBX8 expression was tested by western blot analysis. β-Actin was the loading control. N represents adjacent normal tissues, T represents carcinoma tissues. C. Representative immunohistochemical detection of the CBX8 protein expression in paired esophageal tissues, N represents adjacent normal tissues, T represents carcinoma tissues. Left: hematoxylin-eosin staining (H&E); Right: immunohistochemistry. D. CBX8 expression patterns in normal cell lines and ESCC cell lines.

Article Snippet: For stable CBX8 depletion, the cells were infected with lentivirus (LV) that expressed CBX8-shRNA (Genchem, China) or its negative control at 30-50% confluence.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CBX8, a novel DNA repair protein, promotes tumorigenesis in human esophageal carcinoma

doi:

Figure Lengend Snippet: Association between CBX8 expression and clinicopathologic information of ESCC patients

Article Snippet: For stable CBX8 depletion, the cells were infected with lentivirus (LV) that expressed CBX8-shRNA (Genchem, China) or its negative control at 30-50% confluence.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CBX8, a novel DNA repair protein, promotes tumorigenesis in human esophageal carcinoma

doi:

Figure Lengend Snippet: CBX8 is required for the proliferation of ESCC cells and cell cycle progression. (A) The interference efficiency of two CBX8-siRNAs was tested by both RT-PCR and western blot. (B, C) The relative cell proliferation was measured by MTT (B) and colony formation assays (C) in EC109 and EC9706 cells. (D) Xenograft mouse model. Tumor volume and weight were measured for 30 days after subcutaneous injection. n=5 per group. (E) FACS analysis showed that CBX8-depleted cells accumulated in the G2M phase in EC109. (F) G2M checkpoint-related proteins were detected in EC109 cells. (NC: Negative Control).

Article Snippet: For stable CBX8 depletion, the cells were infected with lentivirus (LV) that expressed CBX8-shRNA (Genchem, China) or its negative control at 30-50% confluence.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Injection, Negative Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CBX8, a novel DNA repair protein, promotes tumorigenesis in human esophageal carcinoma

doi:

Figure Lengend Snippet: CBX8 is related to DNA damage. (A) CBX8-depletion spontaneously induces DNA damage. EC109 cells were transfected with the indicated siRNA for 48 h, exposed to 10 Gy of IR or 0.1 mM H2O2 for 20 min, and subjected to neutral comet analysis. (B) Quantification of tail moments using cells from (A). Tail moments for each condition were calculated on a minimum of 300 cells for each data point. Three independent experiments have been done for each result. *P<0.05, without IR or H2O2 treatment; **P<0.01, IR treatment or H2O2 treatment. (C) A marker for DNA DSBs, γ-H2AX, was tested by western blot both in EC109 and EC9706 cells. (D) EC109 cells transfected with NC or siCBX8 were fixed and immunostained with anti-γ-H2AX antibody.

Article Snippet: For stable CBX8 depletion, the cells were infected with lentivirus (LV) that expressed CBX8-shRNA (Genchem, China) or its negative control at 30-50% confluence.

Techniques: Transfection, Marker, Western Blot

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CBX8, a novel DNA repair protein, promotes tumorigenesis in human esophageal carcinoma

doi:

Figure Lengend Snippet: Depletion of CBX8 decreases resistance to H2O2 or IR. A, B. Both EC109 and HeLa cells were treated with H2O2 or IR. For H2O2 treatment, cells were exposed to 0.1 mM H2O2 for 20 min and then maintained in culture for indicated time points at 37°C, 5% CO2. For IR exposure, cells were exposed to 10 Gy IR and maintained in culture for indicated time points at 37°C, 5% CO2. CBX8 and γ-H2AX were determined by Western blot. C. EC109 cells were transfected with siCBX8 for 48 h, then exposed to 0.1 mM H2O2 for 20 min, and subjected to neutral comet assay at the indicated time points. D. EC109 cells were transfected with siCBX8; exposed to 0.1 mM H2O2 for 20 min; and maintained in culture for indicated time points. CBX8 and γ-H2AX levels were monitored by western blot analysis; β-actin was used as an internal reference. Results indicate that CBX8 depletion increases sensitivity of EC109 cells to H2O2.

Article Snippet: For stable CBX8 depletion, the cells were infected with lentivirus (LV) that expressed CBX8-shRNA (Genchem, China) or its negative control at 30-50% confluence.

Techniques: Western Blot, Transfection, Neutral Comet Assay