Journal:

Article Title: Activation of NF-kB by TMPRSS2/ERG fusion isoforms through Toll-like receptor-4

doi: 10.1158/0008-5472.CAN-10-2210

Figure Lengend Snippet: A. Western blots of extracts of PNT1a cells expressing Type III+72, Type VI+72 or vector control with anti-p65, anti-p65 phospho-Ser536 or β-actin antibodies (left); Similar Western blots oh VCaP cells expressing a fusion gene specific shRNA or vector controls (right). B. Western blot of PNT1a and multiple PCa cell lines with anti-p65, anti-p65 phospho-Ser536 or β-actin antibodies. C and D. Cos7 cells were transfected with NF-κB p65 along with Type III+72 or VI+72 fusion gene expression constructs or control vector (pCMV-Tag2B). TRITC-conjugated goat anti-rabbit antibody was used to detect total p65 (C) or phospho-Ser536 p65 (D) after incubation with anti-p65 or anti-p65 Ser536 antibody. Total positive nuclei per 10 high power fields (200×) is shown as mean +/− standard deviation of 10 fields. Statistically significant differences from controls by t-test are indicated by asterisks (p<.01).

Article Snippet: Immunohistochemistry Immunohistochemistry of VCaP orthotopic tumors was performed using a rabbit polyclonal p65 anti-phospho-Ser536 from Abgent, Inc (AP3178a) as described previously ( 14 ).

Techniques: Western Blot, Expressing, Plasmid Preparation, shRNA, Transfection, Construct, Incubation, Standard Deviation

Journal:

Article Title: Activation of NF-kB by TMPRSS2/ERG fusion isoforms through Toll-like receptor-4

doi: 10.1158/0008-5472.CAN-10-2210

Figure Lengend Snippet: A. Immunohistochemistry using anti-p65 Ser536 antibody was performed using a prostate cancer tissue microarray was performed and quantiated using a 10 point quantitation scale as described in Materials and Methods. Examples of staining with staining indices of 9, 6, 3 and 0 are shown. Original maginification 400×, dash equals 200 microns. B. Kaplan-Meier plot of recurrence free survival following radical prostatectomy of cancers with no staining with anti-p65 Ser536 antibody versus cancers with staining. C. Association of ERG expression with 65 phospho-Ser536 IHC. Cases with the indicated staining indices for p65 phospho-Ser536 by IHC 0, 1–3 (low), 4–6 (moderate) and 7–9 (high) were scored for ERG expression by IHC and the fraction of cases with ERG expression, which is highly correlated with expression of the T/E fusion gene was determined. The distribution of ERG positive cases between groups was significantly different than chance (p<.0001, Chi-square)

Article Snippet: Immunohistochemistry Immunohistochemistry of VCaP orthotopic tumors was performed using a rabbit polyclonal p65 anti-phospho-Ser536 from Abgent, Inc (AP3178a) as described previously ( 14 ).

Techniques: Immunohistochemistry, Microarray, Quantitation Assay, Staining, Expressing

Journal:

Article Title: Activation of NF-kB by TMPRSS2/ERG fusion isoforms through Toll-like receptor-4

doi: 10.1158/0008-5472.CAN-10-2210

Figure Lengend Snippet: A. Expression of TLR4 as determined by quantitative RT-PCR in PNT1a cells stably expressing the VI+72 fusion gene isoform transiently transfected with two plasmids expressing shRNas targeting TLR4 or control plasmid. B. Western blot of protein extracts on transiently transfected PNT1a cells described above using anti-p65 phospho-Ser536 antibody. β-actin is a loading control.

Article Snippet: Immunohistochemistry Immunohistochemistry of VCaP orthotopic tumors was performed using a rabbit polyclonal p65 anti-phospho-Ser536 from Abgent, Inc (AP3178a) as described previously ( 14 ).

Techniques: Expressing, Quantitative RT-PCR, Stable Transfection, Transfection, Plasmid Preparation, Western Blot