<div>Figure 1. Time-based dose response analysis of MT-2 cells to GBA after
24, 48, and 72 h. The MT-2 cell line was treated with various
concentrations of GBA (5, 10, 20, 40, and 80 µM) for 24, 48, and 72 h.
The cell viability was assessed with alamar blue assay. Viability
assessment was carried for at least three times and results are
presented as mean ± SD. MT-2 cells treated with 0.5% DMSO were
considered as relevant control. Statistical significance from control;*P
&lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001, and****P
&lt; 0.0001.</div><div>Figure 2. (A) Viability of MT-2 cells after treatment with various
concentrations of GBA and ATO at 48 h. In each combination, cells
treated with 0.5% DMSO-ATO (with similar dose) were considered as
relevant control. (B)Viability of MT-2 cells significantly decreased
after treatment with 20 µM GBA in combination with 4 µM ATO after 48 h
compared to treatment with each agent alone at equal concentrations.
Viability assessment was carried for at least three times and results
are presented as mean ± SD. Statistical significance from control;*P
&lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001, and****P
&lt; 0.0001.</div><div>Figure 3. Effects of GBA/ATO on apoptotic cell death in MT-2 cell line.
The MT-2 cells were treated with 20 µM GBA and 4 µM ATO alone or in
combination for 48 h and stained with PI. Then the
sub-G<sub>1</sub> population was analyzed by flow cytometry. PI
staining reveals distribution of cells in four major phases of cell
cycle including, sub-G<sub>1</sub>, G<sub>1</sub>, S, and
G2/M. (A) Untreated, (B) 0.5% DMSO-treated, (C) 20 µM GBA-treated, (D)
4 µM ATO-treated, (E) 4 µM 0.5% DMSO-ATO-treated, and (F) 20 µM GBA-4
µM ATO-treated cells.</div><div>Figure 4. Flow cytometric analysis of Mitoxantrone uptake and efflux by
MT-2 cells. P-gp function was assayed by flow cytometry in MT-2 cells
after exposure to 10 µ Mmitoxantrone. As shown by the shift in
fluorescence, GBA decreased mitoxantrone efflux in MT-2 cells (C)
compared to Untreated (A) and DMSO-treated (B) cells.</div><div>Figure 5. The mRNA expression levels of (A) RelA, (B) p53, (C) CDK4, (D)
c-MYC, (E) c-FLIPL, and (F) c-FLIPS in MT-2 cells treated with 20 µM GBA
and 4 µM ATO alone and in combination. Statistical significance from
control, *P &lt; 0.05, **P &lt; 0.01, ***P &lt;
0.001, and****P &lt; 0.0001.</div><div>Figure 6. Pearson’s correlation analysis of the expression of p53 and
c-FLIPS in 6 groups of MT-2 cells treated with 20 µM GBA and/or 4 µM ATO
alone and in combination.</div>