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1.
Figure 2

Figure 2. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib combination sensitises ovarian cancer cells to platinum-based chemotherapeutic agents. (A) OVCAR3 and OVCAR5 cells were treated with I3C, bortezomib, cisplatin or (C) carboplatin at the indicated concentrations for 48 h. Cell viability was measured as described in Materials and Methods. The data shown represent the mean±s.e.m. (n=3). (B) SKOV3, OVCAR8, HEY and CAOV3 cells were treated with I3C, bortezomib and cisplatin as in (A). The data shown represent the mean±s.e.m. (n=3).

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.
2.
Figure 3

Figure 3. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib enhance apoptosis in OVCAR3 and OVCAR5 cells. (A) OVCAR3 and (B) OVCAR5 cells were treated with I3C, bortezomib or their combination for 24 h at the indicated concentrations. The cells were subsequently co-stained with annexin V and PI, and viable (annexin V-negative and PI-negative), early apoptotic (annexin V-positive and PI-negative) and late apoptotic (annexin V-positive and PI-positive) cells were distinguished by flow cytometric analysis. The stacked bar graph represents the mean percentage of viable (grey), early apoptotic (black) and late (white) apoptotic cells from a triplicate experiment.

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.
3.
Figure 6

Figure 6. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib co-treatment inhibit human ovarian tumour xenografts in nude mice. (A) Relative tumour growth of OVCAR5 xenografts treated with vehicle (control), 20 mg kg−1 I3C, 1 mg kg−1 bortezomib or in combination measured from 0 to 53 days post treatment or (B) measured 42, 45, 49 and 53 days post treatment. The data shown represent the mean±s.e.m. (n=8). (C) Representative tumour images of control- (vehicle), I3C- and bortezomib-treated mice pre and post dissection, with corresponding (D) tumour weight 53 days post treatment. The data shown represent the mean±s.e.m. (n=8).

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.
4.
Figure 1

Figure 1. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib exhibit synergistic cytotoxicity in OVCAR3 and OVCAR5 cells. (A) Dose-dependent cytotoxicity of I3C, bortezomib and their combination. OVCAR3 and OVCAR5 cells were treated with I3C and bortezomib at the indicated concentrations for 48 h. Cell viability was measured as described in Materials and Methods. The data shown represent the mean±s.e.m. (n=3). (B) Isobologram analysis of combination I3C with bortezomib used in equipotent concentrations in OVCAR3 and OVCAR5 cells. The line designates the CI where CI=1 (additive effect). Combination index <1 indicates synergism and CI>1 represents antagonism. The combination data points (CI=0.73 for OVCAR3 and CI=0.27 for OVCAR5) calculated by CalcuSyn software indicate synergism at ED50. (C) The CI values of combination I3C and bortezomib at a range of EDs. TheCI values at ED25, ED50, ED75 and ED90 indicate a synergistic interaction between I3C and bortezomib in OVCAR3 and OVCAR5 cells.

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.
5.
Figure 4

Figure 4. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib cause cell cycle arrest in OVCAR3 and OVCAR5 cells. (A) OVCAR3 and (B) OVCAR5 cells were treated with I3C, bortezomib or in combination for 24 h at the indicated concentrations. Cells were fixed and stained with PI followed by flow cytometric analysis for DNA content as described in Materials and Methods. A representative DNA histogram is shown for each condition. The mean percentage of cells from a triplicate experiment is indicated for cells in G1-, S- and G2-M-phase for each condition. (C) OVCAR3 and (D) OVCAR5 cells were treated with I3C, bortezomib or in combination as in (A) and (B), respectively. Whole-cell extracts were isolated and immunoblotted with the indicated antibodies. Actin was used as a loading control.

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.
6.
Figure 5

Figure 5. From: Indole-3-carbinol synergistically sensitises ovarian cancer cells to bortezomib treatment.

Indole-3-carbinol and bortezomib combination inhibits carcinogenesis, reduces chemoresistance, upregulates ER stress markers, deregulates metabolic pathways and causes widespread gene deregulation in OVCAR3 and OVCAR5 cells. OVCAR5 cells were treated with vehicle (mock), 675 μ I3C, 37.5 n bortezomib or in combination for 24 h with subsequent RNA isolation and microarray gene expression analysis. (A) A dendrogram presentation of the data from triplicate experiments indicate that replicate samples cluster together. (B) Venn diagram representing overlap between I3C and bortezomib treatment. Target genes with log-fold changes >0.4 or <−0.4 (P<0.0025) are presented. (C) Quantitative RT–PCR of candidate target genes identified by microarray analysis categorised by function. Target gene validation was also performed in OVCAR3 cells treated with vehicle (mock), 270 μ I3C, 18.8 nM bortezomib or in combination for 24 h. (D) Pleiotropic effect of I3C and bortezomib on multiple biological processes. Representative target genes with log-fold changes >1.5 or <−1.5 (P<0.0025) are shown.

B Taylor-Harding, et al. Br J Cancer. 2012 January 17;106(2):333-343.

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