Journal: Oncotarget

Article Title: Xanthine oxidoreductase is required for genotoxic stress-induced NKG2D ligand expression and gemcitabine-mediated antitumor activity

doi: 10.18632/oncotarget.11042

Figure Lengend Snippet: ( A ) IF staining of XOR. HeLa (a) and HT29 (b) cells were analyzed for XOR expression by IF staining with an anti-XOR antibody. (c) The specificity of the antibody on HeLa cells was confirmed by pre-mixing anti-XOR antibody (1:50 in 200 μl) with purified XOR (2 μg/200 μl). Normal rabbit IgG was included as a negative control (d). ( B – D ) Genotoxic stress does not alter XOR expression. HeLa and HT29 cells were left untreated or treated with 5-FU (10 μM), gemcitabine (2 μM) or radiation (40 Gy) for 24 hr (B) or treated with 5-FU (10 μM) or gemcitabine (2 μM) in HeLa (C) and HT29 cells (D) for the indicated time. Cell lysates were prepared in NP-40 lysis buffer and analyzed for XOR expression by Western blot with a rabbit anti-XOR antiserum.

Article Snippet: XOR expression was detected by IF staining with a rabbit anti-XOR rabbit antibody (Rockland Immunologicals, Inc., Bertsville, PA) (1:100) and examined under a fluorescence microscope.

Techniques: Staining, Expressing, Purification, Negative Control, Lysis, Western Blot

Journal: Oncotarget

Article Title: Xanthine oxidoreductase is required for genotoxic stress-induced NKG2D ligand expression and gemcitabine-mediated antitumor activity

doi: 10.18632/oncotarget.11042

Figure Lengend Snippet: ( A ) Immunofluorescence (IF) staining of XOR. HeLa cells stably transfected with control miRNA (Ctr-miRNA) or XOR-miRNA were analyzed for XOR expression by IF staining with an anti-XOR antibody as described in Figure . Nuclei was stained with DAPI. ( B and C ) XOR knockdown blocks uric acid production and MICA/B expression. Ctr-miRNA and XOR-miRNA-transfected cells were exposed to 5-FU (10 μM), gemcitabine (2 μM), or radiation (40 Gy). Cells were harvested at 24 hr and analyzed for intracellular uric acid levels using a uric acid assay kit (B) or for MICA/B expression by FACS (C). Black line, isotype control; Green line, MICA/B. Inset in (B): XOR expression analyzed by Western blot. Actin was included as a loading control. ( D ) Western blot analysis of XOR expression in Ctr-miRNA- and XOR-miRNA-transfected RCAS-Neu cells. ( E ) XOR knockdown blocks genotoxic stress-induced Rae I expression. Ctr-miRNA- and XOR-miRNA-transfected RCAS-Neucells were exposed to 5-FU (10 μM) or gemcitabine (2 μM) and analyzed for Rae I expression 16 hr later by FACS analysis.

Article Snippet: XOR expression was detected by IF staining with a rabbit anti-XOR rabbit antibody (Rockland Immunologicals, Inc., Bertsville, PA) (1:100) and examined under a fluorescence microscope.

Techniques: Immunofluorescence, Staining, Stable Transfection, Transfection, Control, Expressing, Knockdown, Uric Acid Assay, Western Blot