Analysis of BMI1 and related pathways

Docetaxel, and PTC-209 were purchased from selleckchem (Houston, TX USA). The ChIP-grade anti-BMI1 antibody was obtained from Millipore (Bedford, MA, USA). We used following antibodies: anti-BCL2, anti-BMI1, anti-mitochondrial protein, anti-PCNA, anti-Cyclin D1 (Cell Signaling, Danvers, MA), anti-P16, anti-c-MYC (Abcam, Cambridge, MA) and anti-VEGF (SantCruz, CA). We used following plasmids pTK-TCF-Luc (Upstate Laboratories, Lake Placid, NY); pGL3-MMP2, pGL3-VEGF, pGL3-MYC (Addgene, Cambridge, MA) and pGL3-BMI1 (gifted by Goberdhan Dimri, George Washington University, Washington DC).
Transfections, luciferase reporter activity, Immunostaining, Immunoblot analysis, 3[H]-thymidine incorporation, cell growth, migration and invasion assays were performed as described previously (13 (link), 14 , 16 (link)).

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Ganaie A.A., Beigh F.H., Astone M., Ferrari M.G., Maqbool R., Umbreen S., Parray A.S., Siddique H.R., Hussain T., Murugan P., Morrissey C., Koochekpour S., Deng Y., Konety B.R., Hoeppner L.H, & Saleem M. (2018). BMI1 drives metastasis of prostate cancer in Caucasian and African-American men and is a potential therapeutic target: hypothesis tested in race-specific models. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(24), 6421-6432.

Publication 2018

Docetaxel PTC-209 anti-BCL2 anti-BMI1 anti-PCNA anti-Cyclin D1 anti-P16 anti-c-MYC pGL3-MMP2 pGL3-VEGF

Anti antibody Antibodies Assays Bcl2 Bmi1 Cell Chip Cyclin d1 Docetaxel Immunoblot analysis Luciferase Mitochondrial protein Mmp2 Pcna Pgl3 Plasmids Ptc 209 Thymidine Transfections Vegf

Corresponding Organization : University of Minnesota

Other organizations : Hormel (United States), Hamad Medical Corporation, Aligarh Muslim University, University of Washington, Roswell Park Comprehensive Cancer Center, Cancer Genetics (United States)

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Variable analysis

independent variables

Docetaxel
PTC-209

dependent variables

Cell growth
Migration
Invasion
Luciferase reporter activity
Protein expression (BCL2, BMI1, mitochondrial protein, PCNA, Cyclin D1, P16, c-MYC, VEGF)

control variables

Cell culture conditions
Antibodies used for immunostaining and immunoblot analysis
Plasmids used for transfections and luciferase reporter assays

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