Journal: The American Journal of Pathology

Article Title: Regulation of Apo2L/Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Induced Apoptosis in Thyroid Carcinoma Cells

doi: 10.1016/s0002-9440(10)64220-4

Figure Lengend Snippet: Figure 2. Protective role of FLIP against TRAIL-induced apoptosis in thyroid carcinoma cells. A: TRAIL-resistant SW579-TR cells express higher levels of FLIP, but not Bcl-2, than the parental, TRAIL-sensitive cells. B: Pretreatment of SW579-TR cells with cycloheximide (CHX, 10 g/ml) or bisindolylmale- imide (BIM III, 20 mol/L) restored sensitivity to LZ-TRAIL (300 ng/ml) (no TRAIL, black bars; TRAIL treatment, white bars). Cell survival was quan- tified with the MTT assay. C: CHX and BIM III specifically down-regulated FLIP protein levels in SW579-TR cells after 8 hours of treatment, but not those of cIAP-1, cIAP-2, Bcl-2, or Bcl-xL. Tubulin levels are shown for comparison. D: Indirect confirmation of the anti-apoptotic role of FLIP, SW579-TR cells were sensitized to LZ-TRAIL (300 ng/ml) by FLIP anti-sense oligonucleotides (AS), but not by control oligonucleotides (CO). Cell survival was quantified with the MTT assay.

Article Snippet: Recombinant human TRAIL was obtained from Immunex Corporation (Seattle, WA), in a leucine zipper (LZ) form that promotes and stabilizes the formation of trimers, as previously reported;13 for comparison, several experiments were repeated using the recombinant Apo2L form from Genentech Inc. (South San Francisco, CA).20 The goat polyclonal antibodies for DR4, DR5, DcR1, and mouse monoclonal antibody for tubulin were from Santa Cruz Biotechnologies (Santa Cruz, CA); the anti-DcR2 rabbit polyclonal antibody was from Imgenex (San Diego, CA); cycloheximide, geldanamycin, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was from Sigma Chemical Co. (St Louis, MO); IGF-1, bFGF, EGF, IFN- , and TNF- were from R&D Systems (Minneapolis, MN; bisindolylmaleimide (BIM) III and wortmannin were from Calbiochem (La Jolla, CA); rabbit polyclonal antibody for caspase-10 was from Research Diagnostics Inc. (Flanders, NJ); the IGF-1 receptor neutralizing antibody aIR3 was from Oncogene Research (Cambridge, MA); and the enhanced chemiluminescence (ECL) kit, which includes the peroxidase-labeled anti-mouse and anti-rabbit secondary antibodies, was from Amersham (Arlington Heights, IL).

Techniques: MTT Assay, Comparison, Control

Journal: International journal of molecular medicine

Article Title: Molecular mechanisms of ginsenoside Rp1-mediated growth arrest and apoptosis.

doi: 10.3892/ijmm_00000243

Figure Lengend Snippet: Figure 7. Cleavage of Bid (A), mitochondrial translocation of Bax, and release of cytochrome c from mitochondria (B) in Rp1-treated HeLa cells. The cells were incubated with Rp1 at 40 μM for 48 h. Whole cell fractions (A) or cytosol and mitochondrial fractions (B) were applied to Western blot analysis. Cytochrome oxidase subunit IV (CoxIV) and tubulin were used as cytosolic and mitochondrial loading controls.

Article Snippet: Antibodies against human caspase-3, -8, -9, polyADP-ribose polymerase (PARP), Bad, _________________________________________ Correspondence to: Dr Hiroshi Kimura, Department of Molecular Genetics in Medicine, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan E-mail: kimurah@belle.shiga-med.ac.jp Key words: ginsenoside, apoptosis, cell cycle, Bcl-2 family proteins, mitochondria Bax, Bid, Bcl-2, cytochrome c, cyclin D1 and A, p21, p53, phospho-p53 (Ser15), cytochrome oxidase IV (CoxIV), and tubulin were obtained from Cell Signaling Technology (Beverly, MA), and anti ß-actin antibody was purchased from Sigma (St. Louis, MO).

Techniques: Translocation Assay, Incubation, Western Blot

Journal: Journal of proteome research

Article Title: Proteomic profile identifies dysregulated pathways in Cornelia de Lange syndrome cells with distinct mutations in SMC1A and SMC3 genes.

doi: 10.1021/pr300760p

Figure Lengend Snippet: Figure 6. c-MYC is down-regulated in CdLS cell lines. Western blotting showed that the transcription factor c-Myc is down-regulated in CdLS cell lines, whereas no difference was found for ESR1 and p53 transcription factors, supporting the notion that c-MYC is an important hub for identified proteins. B-Tubulin (B-TUB) was used as loading control.

Article Snippet: To assess equal protein loads, tested membranes were immunostained with anti-β-Tubulin antibody (H-235, Santa Cruz Biotechnology), diluted 1:1000.

Techniques: Western Blot, Control