Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: Expression of p21 protein detected by Western blot analysis. The cervical cancer cells, Z172, SiHa, C33A, and HeLa, were infected with sense p21 or antisense p21 adenovirus at a MOI of 25 or with PBS (mock); after 48 h, protein lysates were isolated and analyzed by immunoblotting with anti-p21 antibody (Pharmingen).

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: Expressing, Western Blot, Infection, Isolation

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: In vitro effect of p21 on cell growth. The growth of these four cervical cancer cell lines was determined by counting the cell number at different time points after infection of cells at a MOI of 25 with sense p21 or antisense p21 adenovirus. The results are the mean of three separate experiments.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: In Vitro, Infection

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: DNA fragmentation assays. Low-molecular-weight DNA was harvested from attached and detached cells infected with sense p21 or antisense p21 adenovirus, and the DNA was analyzed by electrophoresis on a 1.4% agarose gel.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: Molecular Weight, Infection, Electrophoresis, Agarose Gel Electrophoresis

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: In vitro analysis for apoptosis. Cells infected with sense p21 adenovirus, antisense p21 adenovirus, or PBS only (mock) for 2 days were stained with acridine orange and observed by fluorescence microscopy (a and d) HeLa and C33A cells infected with sense p21 adenovirus, respectively; (b and e) HeLa and C33A cells infected with antisense p21 adenovirus, respectively; (c and f) HeLa and C33A cells without virus infection (mock infected), respectively. The apoptotic bodies are indicated by arrowheads in panels a and d.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: In Vitro, Infection, Staining, Fluorescence, Microscopy

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: p21-mediated tumor growth inhibition. HeLa and SiHa cells (2.5 × 106) were separately injected subcutaneously into the flanks of nude mice. When the tumors grew to about 75 mm3, the animals were reanesthetized and 2.5 × 107 PFU of adenovirus encoding sense p21 or antisense p21 or PBS only (mock infection) was injected into the tumor in five nude mice in each cell line group. The volumes of the tumors in five nude mice in each group were measured weekly. The error bars indicate the standard deviations of tumor volumes of five mice.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: Inhibition, Injection, Infection

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: In vivo analysis of apoptosis by an in situ TUNEL assay. In situ end-labeling analysis was performed on paraffin-embedded sections obtained from tumor-bearing nude mice which were injected with sense p21 or antisense p21 adenovirus or PBS only (mock infected) 3 days earlier. SiHa cells were injected with PBS only (a and e), antisense p21 adenovirus (b and f), or sense p21 adenovirus (c, d, g, and h). (a to d) Hematoxylin and eosin stain; (e to h) fluorescent stain of apoptotic cells as shown by the in situ TUNEL assay. Magnifications, ×40 (a, b, c, e, f, and g) and ×300 (d and h); panels d and h are magnifications of panels c and g, respectively.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: In Vivo, In Situ, TUNEL Assay, End Labeling, Injection, Infection, H&E Stain, Staining

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: Expression of p21 in tumor tissues. At 36 h postinfection, the paraffin-embedded sections were obtained from sense p21 virus-treated (a), antisense p21 virus-treated (b) or mock-treated (c) tumors and stained with p21 antibody (Pharmingen) by the immunohistochemistry assay. Hematoxylin was used for counterstaining.

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: Expressing, Staining, Immunohistochemistry

Journal:

Article Title: Adenovirus-Mediated p21 (WAF1/SDII/CIP1) Gene Transfer Induces Apoptosis of Human Cervical Cancer Cell Lines

doi:

Figure Lengend Snippet: Bcl-2 cannot protect against p21-mediated apoptosis in cervical cancer cells. (A) Expression of Bcl-2 protein. Cellular proteins were extracted, then separated by a SDS-PAGE, and subjected to immunoblot analysis with antibody recognizing Bcl-2 (Santa Cruz). Lanes 1 and 2 represent SiHa cells and SiHa/bcl2 cells (which contain the bcl-2 gene). (B) Low-molecular-weight DNA was harvested from attached and detached SiHa/bcl2 cells which were infected with sense p21 (lane 2) or antisense p21 (lane 1) adenovirus and then analyzed by electrophoresis on a 1.4% agarose gel. (C) SiHa cells with or without Bcl-2 protein infected with sense p21 or antisense p21 adenovirus for 2 days were stained with acridine orange and observed with a fluorescence microscope. Panels a and b represent SiHa cells infected with sense p21 and antisense p21 adenovirus, respectively. Panels c and d represent SiHa/bcl2 cells infected with sense p21 and antisense p21 adenovirus, respectively. The apoptotic cells were observed in panels a and c (arrowheads).

Article Snippet: Then the sections were processed as described previously ( 2 ) and incubated with an appropriate dilution of primary antibody against p21 protein (Pharmingen).

Techniques: Expressing, SDS Page, Western Blot, Molecular Weight, Infection, Electrophoresis, Agarose Gel Electrophoresis, Staining, Fluorescence, Microscopy

Journal:

Article Title: Upregulation of CD40 Expression on Endothelial Cells Infected with Human Cytomegalovirus

doi: 10.1128/JVI.76.24.12803-12812.2002

Figure Lengend Snippet: Immunoblot analysis of CD40 in whole-cell lysates. Cell monolayers were infected with HCMV TB40E at an MOI of 1 (inf.) or mock infected (noninfected [n.i.]). Cell lysates were prepared at the indicated times and analyzed by immunoblotting. (A) CD40 protein levels were detected with a polyclonal rabbit serum directed against human CD40. (B) Blots from panel A were stripped and reprobed with a rabbit serum directed against calreticulin. (C) Signal intensities from panels A and B were densitometrically quantified; data represent the ratio of CD40 to calreticulin (y axis) versus time p.i. (x axis). (D) Protein lysates from infected or noninfected cells were treated with N-glycosidase F (+) or left untreated (−) prior to immunoblot analysis with anti-CD40 antibodies. Molecular masses are given in kilodaltons.

Article Snippet: The following monoclonal antibodies or polyclonal sera were used: mouse IgG1-fluorescein isothiocyanate (FITC)-conjugated anti-human CD54 (Calbiochem, La Jolla, Calif.), anti-human major histocompatibility complex class I (MHC-I) (BD PharMingen, San Diego, Calif.), mouse IgG1-FITC anti-human CD40 (Dianova, Hamburg, Germany), mouse IgG1-FITC anti-human CD62E (Calbiochem), antibodies G28.5 and Ro1 (anti-human CD40) (gifts from H. Engelmann, Institute for Immunology, University of Munich, Munich, Germany) ( 39 ), polyclonal rabbit anti-CD40 serum (Santa Cruz Biotechnology), rabbit antiserum against human calreticulin (Dianova), biotin-conjugated rabbit anti-mouse antibodies, and biotinylated swine anti-rabbit antibodies (Dako, Hamburg, Germany).

Techniques: Western Blot, Infection