Journal: Molecular oncology

Article Title: Increased extracellular pressure stimulates tumor proliferation by a mechanosensitive calcium channel and PKC-β

doi: 10.1016/j.molonc.2014.10.008

Figure Lengend Snippet: Pressure induces proliferation through Cav3.3 and NF‐kB. (7A) SW620 cells were incubated under ambient or 40 mmHg increased pressure for 24 h in the presence of an NF‐kB inhibitor SN50 (12 μM) or its control peptide, an NF‐kB nuclear decoy (25 μM) or its inactive control, an NF‐kB inhibitor TCH 021 (1 μM), or a rac1 inhibitor (NSC23766 30 μM) before MTT assay. (7B) S‐phase fraction was measured in previously serum‐starved SW620 cells via flow cytometry. Before measurement, cells were incubated under ambient or 40 mmHg increased pressure for 24 h in the presence of an NF‐kB inhibitor SN50 (12 μM) or its control peptide, an NF‐kB nuclear decoy (25 μM) or its inactive control, an NF‐kB inhibitor TCH 021 (1 μM). (7C) SW620 cells were transfected with siRNA targeting Cav3.3 or with a non‐targeting control for 48hr and exposed to ambient or 15 mmHg for 24hr before being lysed and immunoblotted with cyclin D1 antibodies and normalized to GAPDH. (*p < 0.05 vs. paired controls).

Article Snippet: Mouse monoclonal antibodies to cyclin D1, pIkB (ser 32/36, Cell Signaling), and actin (Sigma–Aldrich) were also used, with a horseradish peroxidase conjugated anti‐mouse secondary antibody.

Techniques: Incubation, Control, MTT Assay, Flow Cytometry, Transfection

Journal: Molecular oncology

Article Title: Increased extracellular pressure stimulates tumor proliferation by a mechanosensitive calcium channel and PKC-β

doi: 10.1016/j.molonc.2014.10.008

Figure Lengend Snippet: Immunohistochemical staining of colorectal carcinomas. Staining of colorectal carcinomas was representative of that observed in colorectal, head and neck and lung tumors. To obtain a full view of the sample, a panel of overlapping photographs were taken and placed together. Dashed lines represent zones of overlap within the image. Overall panel images were taken at 40× magnification and higher power images within each zone at 200×. NF‐kB (8A), phospho‐IkB (8B), and cyclin D1 (8C) immunoreactivity and mitotic index (8D) are increased in tumor centers vs. peripheries. Non‐malignant tissue showed minimal staining and few mitotic figures.

Article Snippet: Mouse monoclonal antibodies to cyclin D1, pIkB (ser 32/36, Cell Signaling), and actin (Sigma–Aldrich) were also used, with a horseradish peroxidase conjugated anti‐mouse secondary antibody.

Techniques: Immunohistochemical staining, Staining

Journal: Molecular oncology

Article Title: Increased extracellular pressure stimulates tumor proliferation by a mechanosensitive calcium channel and PKC-β

doi: 10.1016/j.molonc.2014.10.008

Figure Lengend Snippet: Immunohistochemical scores.

Article Snippet: Mouse monoclonal antibodies to cyclin D1, pIkB (ser 32/36, Cell Signaling), and actin (Sigma–Aldrich) were also used, with a horseradish peroxidase conjugated anti‐mouse secondary antibody.

Techniques: Immunohistochemical staining