bmi1 sirna  (Santa Cruz Biotechnology)

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing of Bmi-1 Gene Enhances Chemotherapy Sensitivity in Human Glioblastoma Cells

doi: 10.12659/MSM.893754

Figure Lengend Snippet: The expression of BMI1 mRNA in brain glioma tissue by real time-PCR analysis. The expression of BMI1 was higher in brain glioma tissue compared to peri-cancerous tissue ( P <0.05). * P <0.05 vs. peri-cancerous tissue.

Article Snippet: Then, 1 mg of siRNA-BMI1 or siRNA-control (Genepharma, China) was mixed with Enhancer R, followed by mixing with 4 μl TransMessenger, and then 900 μl Serum-free medium was added for incubating the non-transfected U251 cells.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing of Bmi-1 Gene Enhances Chemotherapy Sensitivity in Human Glioblastoma Cells

doi: 10.12659/MSM.893754

Figure Lengend Snippet: Correlation between BMI1 mRNA expression in brain glioma tissue and pathological differentiation.

Article Snippet: Then, 1 mg of siRNA-BMI1 or siRNA-control (Genepharma, China) was mixed with Enhancer R, followed by mixing with 4 μl TransMessenger, and then 900 μl Serum-free medium was added for incubating the non-transfected U251 cells.

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing of Bmi-1 Gene Enhances Chemotherapy Sensitivity in Human Glioblastoma Cells

doi: 10.12659/MSM.893754

Figure Lengend Snippet: ( A ) Representative image of the expression level of BMI1 protein. GAPDH was used as a reference control. ( B ) quantitative analysis of the relative protein levels of BMI1 normalized to those of GAPDH is shown. Data are mean ±SD of 3 independent experiments. * P <0.05. # P<0.05

Article Snippet: Then, 1 mg of siRNA-BMI1 or siRNA-control (Genepharma, China) was mixed with Enhancer R, followed by mixing with 4 μl TransMessenger, and then 900 μl Serum-free medium was added for incubating the non-transfected U251 cells.

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing of Bmi-1 Gene Enhances Chemotherapy Sensitivity in Human Glioblastoma Cells

doi: 10.12659/MSM.893754

Figure Lengend Snippet: ( A ) Impact of BMI1 gene silencing on the cell viability of U251 cells. ( B ) Impact of BMI1 gene silencing on the apoptosis rate of U251 cells. *, # P <0.05 vs. control U251 cells.

Article Snippet: Then, 1 mg of siRNA-BMI1 or siRNA-control (Genepharma, China) was mixed with Enhancer R, followed by mixing with 4 μl TransMessenger, and then 900 μl Serum-free medium was added for incubating the non-transfected U251 cells.

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Silencing of Bmi-1 Gene Enhances Chemotherapy Sensitivity in Human Glioblastoma Cells

doi: 10.12659/MSM.893754

Figure Lengend Snippet: ( A ) Impact of different concentrations of DDP on the apoptosis rate of U251 cells and BMI1-slienced U251 cells. ( B ) Impact of different concentrations of DDP on the cell viability of U251 cells and BMI1-slienced U251 cells. *, # , & , % , ^, @ P <0.05 vs. corresponding control U251 cells.

Article Snippet: Then, 1 mg of siRNA-BMI1 or siRNA-control (Genepharma, China) was mixed with Enhancer R, followed by mixing with 4 μl TransMessenger, and then 900 μl Serum-free medium was added for incubating the non-transfected U251 cells.

Techniques:

Journal: Cell Death & Disease

Article Title: BMI1 nuclear location is critical for RAD51-dependent response to replication stress and drives chemoresistance in breast cancer stem cells

doi: 10.1038/s41419-022-04538-w

Figure Lengend Snippet: A Representative images of BMI1 staining (green bodies). Nuclei are counterstained with DAPi (blue staining). BMI1 protein can accumulate in CAP bodies (left panel) or has uniform punctate distribution (PcG bodies, right panel). Scale bar: 5 μm. B Stacked bar plot representing the proportion of CAP and PcG bodies in ALDH br and ALDH neg SUM159 cells sorted according to their cell-cycle phase (G0/G1, S, and G2/M). C , D Representative images (left panels) of BMI1 costaining (green foci) with γH2AX (red foci) for cells harboring CAP bodies ( C ) or PcG bodies ( D ). Nuclei are counterstained with DAPi (blue staining). The red lines correspond to the line scans. Pearson’s coefficient evaluated the amount of colocalization. Scale bar: 5 μm. On the right panels, line-scan profiles of the relative intensity of BMI1 and γH2AX fluorescent signals. E Bar plots representing the proportion of replicative ALDH br and ALDH neg SUM159 cells with BMI1/γH2AX colocalization. F , G Representative images (left panels) of BMI1 costaining (green foci) with RAD51 (red foci) for cells harboring CAP bodies ( F ) or PcG bodies ( G ). Nuclei are counterstained with DAPi (blue staining). The red lines correspond to the line scans. Pearson’s coefficient evaluated the amount of colocalization. Scale bar: 5 μm. On the right panels, line-scan profiles of the relative intensity of BMI1 and RAD51 fluorescent signals. H Bar plots representing the proportion of replicative ALDH br and ALDH neg SUM159 cells with BMI1/RAD51 colocalization. I Representative images of BMI1 staining (green bodies) in ALDH br and ALDH neg SUM159 cells treated with 5-aza or untreated (CTRL). Nuclei are counterstained with DAPi (blue staining). J Stacked bar plot representing the proportion of CAP and PcG bodies in replicative ALDH br and ALDH neg SUM159 cells treated with 5-aza or untreated. K Bar plots representing the proportion of γH2AX-positive cells for each cell subpopulation (replicative ALDH br and ALDH neg SUM159 cells) following 5-aza treatment or in untreated conditions (CTRL). Statistical test used is Student’s t -test. Data represent mean ± SD.

Article Snippet: We used siRNA gene silencing to specially target and knockdown BMI1 gene. siRNA (6442 S, Cell Signaling) was lipoplexed with Lipofectamine RNAiMAX (Life Technologies) in a 96-well culture plate or in a 100 mm tissue-culture dish.

Techniques: Staining