Journal: Cancer Prevention Research

Article Title: Garcinol, a Polyisoprenylated Benzophenone Modulates Multiple Proinflammatory Signaling Cascades Leading to the Suppression of Growth and Survival of Head and Neck Carcinoma

doi: 10.1158/1940-6207.capr-13-0070

Figure Lengend Snippet: Figure 5. Garcinol suppresses STAT3- and NF-kB–regulated gene products involved in proliferation, survival, and angiogenesis and activates caspase-3. The same blots were stripped and reprobed with b-actin antibody to verify equal protein loading (A and B). A, CAL27 cells (5 105/mL) were treated with 25 mmol/L garcinol for indicated time; whole-cell extracts were prepared, separated by SDS–PAGE, and subjected to Western blotting against cyclin D1, Bcl-2, Bcl-xL, Mcl-1, survivin, and VEGF antibodies. B, the same whole-cell extracts were analyzed by Western blotting using caspase-3 and PARP antibodies. C, CAL27 cells (1 106/mL) were treated with 25 mmol/L garcinol for indicated time intervals, after which RNA samples were extracted. One microgram portions of the respective RNA extracts were subjected to reverse transcription to generate corresponding cDNA. Real-time PCR was conducted to measure the relative quantities of mRNA. Each RT product was targeted against cyclin D1, Mcl-1, Bcl-xL, and VEGF TaqMan probes, with 18S as endogenous control for measurement of equal loading of RNA samples. The results were analyzed using Sequence Detection Software version 1.3 provided by Applied Biosystems. Relative gene expression was obtained after normalization with endogenous Hu18S and determination of the difference in Ct between treated and untreated cells using 2DDCt method.

Article Snippet: Antibodies against p65, IkBa, cyclin D1, Bcl2, Bcl-xL, Mcl-1, survivin, VEGF, caspase-3, PARP, goat antirabbit–horseradish peroxidase (HRP) conjugate, Ki-67, goat anti-mouse HRP, and annexin V–fluorescein isothiocyanate (FITC) assay kit were obtained from Santa Cruz Biotechnology.

Techniques: SDS Page, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Sequencing, Software, Gene Expression

Journal: Cancer Research

Article Title: BCR-ABL Activity Is Critical for the Immunogenicity of Chronic Myelogenous Leukemia Cells

doi: 10.1158/0008-5472.can-07-0302

Figure Lengend Snippet: Figure 4. Expression of antigens in K-562 and K-562R cells in response to imatinib treatment. Representative results of three independent experiments. A, Western blot analysis showing down-regulation of the tumor antigens survivin and adipophilin as well as the antiapoptotic proteins Bcl-2 and Bcl-xL in K-562 cells after imatinib treatment. B, real-time PCR analysis showing down-regulation of the tumor antigen WT-1 in K-562 cells. Columns, mean; bars, SE. C, real-time PCR showing down-regulation of the tumor antigen hTERT in K-562 cells. Columns, mean; bars, SE.

Article Snippet: The blot was probed with antibodies to human survivin (R&D Systems), adipophilin (Progen Biotechnik), Bcl-2 (Santa Cruz Biotechnology), Bcl-xL (BD Biosciences), x-linked inhibitor of apoptosis protein (xIAP; BD Biosciences), PTEN (BD Biosciences), phosphorylated mammalian target of rapamycin (p-mTOR; Cell Signaling Technology), and mTOR (Cell Signaling Technology).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction