bcl 2 coding sequence  (Addgene inc)

Journal: PLoS ONE

Article Title: AS1411-Induced Growth Inhibition of Glioma Cells by Up-Regulation of p53 and Down-Regulation of Bcl-2 and Akt1 via Nucleolin

doi: 10.1371/journal.pone.0167094

Figure Lengend Snippet: (a) Cell cycle arrest induced by AS1411 in U87, U251, SHG44 and NHA cells. 48 h after treatment with AS1411 (5μM), cells were collected and stained with propidium iodide (PI); DNA content was determined by flow cytometry. This assay was performed in triplicate. CRO (5μM) was used as negative control. (b) Histograms showing the percentage of glioma cells and NHA in G0-G1, S, and G2-M phases in four independent experiments. (c) The G2/M cell cycle related protein cyclin A1, cyclin B1 and cyclin D1 was detected by immunoblotting after treatment with AS1411 (5μM) for 48 h, CRO (5μM) was used as negative control, β-actin was used as loading control. (d) Histograms showing the percentage of glioma cells in G0-G1, S, and G2-M phases after the NCL overexpression. Error bars indicate ± s.d. **P<0.01, two-tailed student’s t-test.

Article Snippet: For the gene overexpression, pCEP4 Bcl-2 (Plasmid #16461), pCDH-puro-myr-HA-Akt1 (Plasmid #46969) and GFP-Nucleolin (Plasmid #28176) were obtained from the Addgene (Cambridge, MA).

Techniques: Staining, Flow Cytometry, Negative Control, Western Blot, Control, Over Expression, Two Tailed Test

Journal: PLoS ONE

Article Title: AS1411-Induced Growth Inhibition of Glioma Cells by Up-Regulation of p53 and Down-Regulation of Bcl-2 and Akt1 via Nucleolin

doi: 10.1371/journal.pone.0167094

Figure Lengend Snippet: (a) Apoptotic cell death induced by AS1411 in U87, U251, SHG44 and NHA cells. 48h after treatment, cells were collected and stained with PI and Annexin V–FITC, Annexin V-positive/PI-negative cells were measured by flow cytometry. This experiment was repeated three times. (b) Histograms showing the percentage of cells in apoptosis. U87, U251, SHG44 and NHA cells in four independent experiments. Annexin V-positive cells were considered as apoptotic cells. (c) Histograms showing the percentage of glioma cells apoptosis after the Bcl-2 overexpression and p53 siRNA silencing. Error bars indicate ± s.d. **P<0.01, two-tailed student’s t-test.

Article Snippet: For the gene overexpression, pCEP4 Bcl-2 (Plasmid #16461), pCDH-puro-myr-HA-Akt1 (Plasmid #46969) and GFP-Nucleolin (Plasmid #28176) were obtained from the Addgene (Cambridge, MA).

Techniques: Staining, Flow Cytometry, Over Expression, Two Tailed Test

Journal: PLoS ONE

Article Title: AS1411-Induced Growth Inhibition of Glioma Cells by Up-Regulation of p53 and Down-Regulation of Bcl-2 and Akt1 via Nucleolin

doi: 10.1371/journal.pone.0167094

Figure Lengend Snippet: (a) AS1411 inhibited the migration of glioma cells in vitro. The migration capabilities of U87, U251, SHG44 and NHA cells were assessed after pretreatment with AS1411 of 0 to 10 μM and CRO of 10μM. Akt1 overexpression antagonized AS1411 induced migration inhibition. (b) AS1411 inhibited the invasion of glioma cells in vitro. The invasive capabilities of U87, U251, SHG44 and NHA were assessed after pretreatment with AS1411 of 0 to 10μM and CRO of 10μM. Akt1 overexpression antagonized AS1411 induced invasion inhibition. Error bars indicate s.d. *P<0.05, **P<0.01, two-tailed Student’s t -test.

Article Snippet: For the gene overexpression, pCEP4 Bcl-2 (Plasmid #16461), pCDH-puro-myr-HA-Akt1 (Plasmid #46969) and GFP-Nucleolin (Plasmid #28176) were obtained from the Addgene (Cambridge, MA).

Techniques: Migration, In Vitro, Over Expression, Inhibition, Two Tailed Test