Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: Expression of galectin-1 in HeLa and C33A cells. ( a ) Cells transfected with GFP only, GFP plus galectin-1, or shRNA were compared with cells without a vector. Lane 1: wild type; lane 2: empty vector; lane 3: shRNA; lane 4: GFP; lane 5: GFP+galectin-1; lane 6: wild type; lane 7: GFP; and lane 8: GFP+galectin-1. ( b ) Interaction between endogenous galectin-1 and GFP-galectin-1 fusion protein in C33A and HeLa cells. IP was performed for GFP or galectin-1, and then immunoblotting was performed for galectin-1 in both cells transfected with fusion protein. A band (43 kDa) on each lane indicates fusion protein expression formation. A band (14 kDa) on Lane 1 indicates heterodimer formation of endogenous galectin-1 and fusion protein. ( c ) Distribution of galectin-1 (red color) and GFP-galectin-1 fusion protein (green color) in HeLa cells transfected with GFP-galectin-1 fusion protein

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Transfection, shRNA, Plasmid Preparation, Western Blot

Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: Galectin-1 inhibits radiation-induced cell death. Clonogenic survival curves were compared between ( a ) galectin-1 knockdown and scrambled RNA in HeLa cells 48 h after transfection ( b ) galectin-1 cDNA and GFP vectors in HeLa cells ( c ) galectin-1 knockdown and scrambled RNA in C33A cells 48 h after transfection ( d ) galectin-1 cDNA and GFP vectors in C33A cells. Cells were irradiated and clonogenic assay performed 12 to 14 days following irradiation. Clonogenic assays were performed during three independent experiments at each paired condition (with and without galectin-1 modulation). The error bar represents the standard error of mean: * P <0.05, ** P <0.01, and *** P <0.001

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Transfection, Irradiation, Clonogenic Assay

Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: The interaction between H-Ras and galectin-1 is more intense than that between K-Ras and galectin-1 in C33A and HeLa cells. An in situ proximity ligation assay (PLA) and IP were performed for the interaction between galectin-1 and Ras. ( a ) PLA for galectin-1 and H-Ras in C33A cells with or without galectin-1 overexpression. ( b ) PLA for galectin-1 and K-Ras in C33A cells with or without galectin-1 overexpression. ( c ) PLA for galectin-1 and H-Ras in HeLa cells with or without galectin-1 knockdown. ( d ) PLA for galectin-1 and K-Ras in HeLa cells with or without galectin-1 knockdown. ( e ) PLA for galectin-1 and H-Ras in HeLa cells with or without the galectin-1 inhibitor anginex (10 μ M), which was administered for 24 h before cell fixation. ( f ) IP for galectin-1 and Ras in C33A and HeLa cells transfected with GFP plus galectin-1. IP was performed for GFP, H-Ras, and galectin-1 before immunoblotting for galectin-1, H-Ras, and K-Ras in both cells

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: In Situ, Proximity Ligation Assay, Over Expression, Knockdown, Transfection, Western Blot

Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: Effects of galectin-1 on the Ras-transmitted downstream signals. Scrambled RNA or galectin-1 shRNA were transfected in HeLa cells. GFP or galectin-1 cDNA were transfected in C33A cells. The cells were irradiated with 6 Gy and harvested 5 or 10 min later. Expressions of activated H-Ras, p-Raf-1, and p-ERK using western blots in HeLa and C33A cells were compared with or without galectin-1 modulation

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: shRNA, Transfection, Irradiation, Western Blot

Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: Galectin-1 enhanced the repair of DNA damage. GFP or galectin-1 plus GFP were transfected into C33A cells. Stable clones of scrambled RNA or galectin-1 shRNA were transfected into HeLa cells. Cells were harvested immediately, 4 or 24 h following 6-Gy irradiation. Comet assay was performed. ( a ) A longer tail was noted in galectin-1 shRNA compared with scrambled RNA HeLa cells 24 h following irradiation. ( b ) A shorter tail was noted in galectin-1-overexpressed cells compared with GFP-transfected C33A cells 24 h following irradiation. ( c ) Western blots were performed after harvesting the cells. Greater γ -H2AX expression was noted in galectin-1-knocked down HeLa cells and GFP-transfected C33A cells 4 h and 24 h following irradiation, respectively

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: Transfection, Clone Assay, shRNA, Irradiation, Single Cell Gel Electrophoresis, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: A novel radioresistant mechanism of galectin-1 mediated by H-Ras-dependent pathways in cervical cancer cells

doi: 10.1038/cddis.2011.120

Figure Lengend Snippet: Plating efficiency (%) of clonogenic assay

Article Snippet: 293T cells (CRL-11268), HeLa cells (CCL-2), and C33A (HTB-31) were purchased from the American Type Culture Collection (ATCC).

Techniques: Mutagenesis