Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 1. CNL-MS3 data-dependent analysis of products of reactions of ctDNA with illudin S in the presence of PTGR1 and NADPH. Data are normalized to the base peak in each spectra, and the relative intensity is given as a normalized level (NL). (A) Full-scan MS (NL 1.55E6). (B) Full- scan MS of m/z 384 (NL 9.88E5). (C) Triggered MS2 of m/z 384.44 (NL 1.25E6). (D) Mass spectra of the peak in panel C with major fragment m/z 336 and minor fragments m/z 249 and 201 (inset).

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques:

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 3. Cell viability of drug-treated SW-480 and PTGR1-480 cells (48 h of treatment time). (A) AF-treated cells (n = 4). (B) Illudin S- treated cells (n = 3).

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques:

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 2. Western blot analysis of PTGR1 levels in SW-480 and PTGR1-480 cells. Lanes 1, 6, and 10, marker; lanes 2−4, SW-480 cells; lane 5, SW-837 cells (positive control); and lanes 7−9, PTGR1-480 cells. All lanes are loaded with 70 μg of protein, except lane 4 (62 μg).

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques: Western Blot, Marker, Positive Control

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 5. Time-dependent formation of AF- or illudin S-derived Ade adducts in drug-treated SW-480 and PTGR1-480 cells. (A) 3-AF−Ade, cells treated with 500 nM AF (n = 3). (B) Illudin S−Ade, cells treated with 20 nM illudin S (n = 3). Asterisks denote statistically significant differences between levels in SW-480 vs PTGR1-480 cells (p < 0.05).

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques: Derivative Assay

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 4. Concentration-dependent formation of Ade adducts in drug- treated SW-480 and PTGR1-480 cells (24 h of treatment time). (A) 3- AF−Ade DNA adducts (n = 4). (B) Illudin S−Ade DNA adducts (n = 3). Asterisks denote statistically significant differences between levels in SW-480 vs PTGR1-480 cells (p < 0.05).

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques: Concentration Assay

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 6. Ade adduct formation by AF and illudin S in SW-480 and PTGR1-480 cells as a function of fold-IC50.

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques:

Journal: Chemical research in toxicology

Article Title: Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities.

doi: 10.1021/tx300430r

Figure Lengend Snippet: Figure 7. Model illustrating the steps required to form a DNA adduct from a bioactivated drug. This model is used to explain how different levels of PTGR1 do not influence illudin S−DNA adduct levels.

Article Snippet: Stable transfection of 2 × 10 5 SW-480 cells was performed in a six-well plate with 1 μg of Myc-DDK tagged PTGR1 cDNA (OriGene Technologies, Rockville, MD) with Lipofectamine reagents (Invitrogen, Inc.) and selected with Geneticin (Invitrogen, Inc.).

Techniques: