Journal: PLoS ONE

Article Title: Induction of interferon-β and interferon signaling by TRAIL and Smac mimetics via caspase-8 in breast cancer cells

doi: 10.1371/journal.pone.0248175

Figure Lengend Snippet: MCF-7 cells were treated with LCL161 (10 μM) and/or TRAIL (100 ng/mL) for indicated time periods and total cell lysates were analyzed by immunoblotting for cIAP1 (A) the NFKB2 gene product p100 and p52 (B) or phosphorylated and total STAT1 (D). Intensity of bands in B and D were quantified (C and E) and normalized to LCL161+TRAIL (C) or TRAIL (E) treatment for 48 h. Protein levels of cIAP1 (F), p100 and p52 (G), or phosphorylated and total STAT1 (H) were measured in CAMA-1 cells that had been treated with LCL161 and/or TRAIL for 8 h in the absence or presence of zVAD-FMK (20 μM). Figures are representative of three independent experiments. Data in C and E indicate individual data points with bars representing the mean and error bars representing SEM, n = 3. * denotes p<0.05, ** p<0.01, *** p<0.001 using ANOVA followed by Tukey’s honest significance test.

Article Snippet: This was followed by incubation with primary antibodies against phosphorylated STAT1 (1:400, #9167), total STAT1 (1:400, #9172), FADD (1:400, #27825), caspase-8 (1:300, #9746), and FLIP (1:400, #56343) all from Cell Signaling Technology; p100/p52 (1:300, ab131539) and IFNAR1 (1:400, ab124764) from Abcam; cIAP1 (1:200, AF8181, R&D Systems); actin (1:2,000, #0869100, MP Biomedicals); and RIP1 (1:500, #610458, BD Biosciences).

Techniques: Western Blot

Journal: Oncotarget

Article Title: The BIRC6 gene as a novel target for therapy of prostate cancer: dual targeting of inhibitors of apoptosis

doi:

Figure Lengend Snippet: (A) Correlation of immunohistochemical staining intensity of BIRC6 and clinical (T) stages of prostate cancer (mean staining intensity ± S.E.M.). (B-D) Correlation of BIRC6 immunohistochemical staining intensity with the absence and presence of poor prognostic factors, such as recurrence of PSA, lymph node metastasis and prostatic capsule invasion. The statistical significance of positive trends was determined by the Chi square test for trend. (E) Representative images of correlated expressions between BIRC6 and survivin, XIAP and cIAP1. 20x magnification, scale bar, 100 μm.

Article Snippet: Immunohistochemical staining using rabbit polyclonal antibody against BIRC6 (NB110-40730, Novus Biological, 1:50), rabbit monoclonal antibody against Survivin (#2808, Cell Signaling, 1:50), monoclonal antibody against cIAP1 (MAB8181, R & D Systems, 1:200) and rabbit polyclonal antibody against XIAP (#sc-11426, Santa Cruz, 1:25) was conducted using a Ventana autostainer (model Discover XT; Ventana Medical System, Tucson, AZ) with an enzyme-labelled biotin-streptavidin system and a solvent-resistant DAB Map kit (Ventana).

Techniques: Immunohistochemical staining, Staining

Journal: Oncotarget

Article Title: The BIRC6 gene as a novel target for therapy of prostate cancer: dual targeting of inhibitors of apoptosis

doi:

Figure Lengend Snippet: (A-B) Western blotting showing protein levels of BIRC6, cIAP1 and survivin in two CRPC cell lines (A) PC-3 cells and (B) C4-2 cells transfected with Mock or increasing dosages of scrambled ASO (Scrb), dASOs 6w2 and 6w5 for 72 hr. (C) Comparison of dual IAP targeting and single IAP-targeting. Cell viability of PC-3 cells transfected with dASOs 6w2, 6w5 and siRNA-targeting BIRC6, cIAP1 or survivin, was determined by MTS assay at 72 hr after transfection. Cell viabilities of ASO- and siRNA-treated cells were normalized with corresponding Scrb ASO and siRNA controls. Error bars represent mean percentage cell viability ± S.D. Western blotting of 3 IAPs showing comparable amounts of reduced protein expression obtained with dASO and siRNA single IAP-targeting. (D) Proliferation of PC-3 cells transfected with mock, Scrb ASO, dASOs 6w2 and 6w5. Error bars represent mean cell number ± S.D. (E) MTS viability assay of C4-2 cells treated with dASOs.

Article Snippet: Immunohistochemical staining using rabbit polyclonal antibody against BIRC6 (NB110-40730, Novus Biological, 1:50), rabbit monoclonal antibody against Survivin (#2808, Cell Signaling, 1:50), monoclonal antibody against cIAP1 (MAB8181, R & D Systems, 1:200) and rabbit polyclonal antibody against XIAP (#sc-11426, Santa Cruz, 1:25) was conducted using a Ventana autostainer (model Discover XT; Ventana Medical System, Tucson, AZ) with an enzyme-labelled biotin-streptavidin system and a solvent-resistant DAB Map kit (Ventana).

Techniques: Western Blot, Transfection, Comparison, MTS Assay, Expressing, Viability Assay