Journal: Cell Death & Disease

Article Title: The novel c-Met inhibitor cabozantinib overcomes gemcitabine resistance and stem cell signaling in pancreatic cancer

doi: 10.1038/cddis.2013.158

Figure Lengend Snippet: Long-term treatment with cabozantinib changes the level of anti- and pro-apoptotic proteins. ( a ) Proteins were isolated from parental BxPc-3 cells and the derived subclone XL-7. Binding of proteins to antibodies spotted in duplicate to the membrane of a Human Apoptosis Array was detected using biotinylated secondary antibodies, streptavidin-HRP and chemiluminescence. The pixel density was quantified using ImageJ software and normalized to the mean pixel intensity of the reference spots A1, A12 and E1 on the membrane. Spot D12 is the PBS-negative control. ( b ) mRNA from parental BxPc-3 cells and derived subclones XL-1 to XL-7 was harvested and expression of Bcl-2 and Survivin were examined by qRT-PCR. ( c ) Likewise, expression of anti-apoptotic (Bcl-2, Survivin, XIAP, cIAP2) and pro-apoptotic (p53, Bim) proteins were examined by western blot analysis

Article Snippet: The following antibodies were used: Mouse mAb against human XIAP (BD Biosciences, San Jose, CA, USA), rabbit mAb against human c-Met, SOX2, p53 (Cell Signaling, Danvers, MA, USA), Ki67 (Epitomics, Burlingame, CA, USA), rabbit pAb against human Bcl-2 (Cell Signaling), Bim (Abcam, Cambridge, UK), cIAP2 (Epitomics), survivin and p-Met (R&D Systems, Minneapolis, MN, USA).

Techniques: Isolation, Derivative Assay, Binding Assay, Software, Negative Control, Expressing, Quantitative RT-PCR, Western Blot

Journal:

Article Title: Herpes Simplex Virus Type 2 Induces Rapid Cell Death and Functional Impairment of Murine Dendritic Cells In Vitro

doi: 10.1128/JVI.77.20.11139-11149.2003

Figure Lengend Snippet: Subpopulation of HSV-2-infected BMDC exhibit DNA breaks shortly after infection. BMDC were infected with wt HSV-2 at an MOI of 4 or left untreated, incubated for up to 12 h p.i., and then double stained for HSV ICP8 (anti-ICP8 MAb 383 plus FITC-conjugated goat anti-mouse MAb) and for apoptosis (biotin-labeled dCTP followed by avidin-Texas Red). Shown are immunofluorescence microscope images of the same optical section of a population of HSV-2-infected DC collected 2 h p.i. to demonstrate staining for ICP8 (green) (A), nick end-labeled DNA breaks (TUNEL) (red) (B), and a double-labeled, HSV-2-infected, apoptotic BMDC (yellow). Arrow in panels A, B, and C indicates positive staining of an apoptotic, HSV-2-infected BMDC. Included were double-stained uninfected BMDC as a negative control (D) and DNase-treated uninfected DC stained with biotin-dCTP-avidin-Texas red as a positive control (E) for the TUNEL assay. Magnification, ×40.

Article Snippet: Cytospin slides were prepared and fixed as described above, blocked with 20% goat serum (Jackson ImmunoResearch), and then stained with antibodies against HSV ICP4 (M612446; BioClone), ICP8 (383; a kind gift from David Knipe, Harvard Medical School, Boston, Mass.), or HSV-2 glycoprotein D (gD) (910; Biodesign International, Saco, Maine), followed by FITC-conjugated goat anti-mouse MAb (Jackson ImmunoResearch).

Techniques: Infection, Incubation, Staining, Labeling, Avidin-Biotin Assay, Immunofluorescence, Microscopy, TUNEL Assay, Negative Control, Positive Control