Figure 5.

Efficacy of ADI-PEG20, DTX, and GEM combination across cancers. A, Protein expression of ASS1 in sarcoma cell lines and the corresponding LTAT cell lines [HTB-93 (WT and LTAT), HT-1080 (WT and LTAT), SK-UT-1 (WT and LTAT), and SK-LMS-1 (WT and LTAT)], a melanoma cell line [SK-MEL-2 (WT and LTAT)], and pancreatic cell lines [As-PC-1 (WT and LTAT), MiaPaCa-2 (WT and LTAT), and HPAC (WT and LTAT)]. Protein extracts were analyzed by a WES automated blotting system. Band density differences were expressed as relative densitometry normalized to the total protein in each capillary. B, Functional assays using 5-proparglyamino-cytidine-5-carboxyfluorescein probe. Probe uptake was measured at 492 nmol/L and normalized to cell count within the well (NucLight Red). Sarcoma, melanoma, and pancreatic WT and LTAT cells were treated with combo or combo (+c-Myc inhibitor) for various time points. C, Sarcoma, melanoma, and pancreatic cells were treated with combo or combo (+c-Myc inhibitor) and real-time increase in expression of hENT1 was quantified by live-cell immunocytochemistry on the IncuCyte system. D, Cell death activity was measured using YOYO-1 nucleic acid dye on the IncuCyte system. Green counts/well were measured and normalized to total nuclear count within the well (NucLight Red). Sarcoma, melanoma, and pancreatic cell lines were treated with c-Myc inhibitor, combo or combo (+c-Myc inhibitor). Data are from 3 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.