Skip to main content
. Author manuscript; available in PMC: 2012 Jul 15.
Published in final edited form as: Cancer Res. 2011 May 27;71(14):4955–4967. doi: 10.1158/0008-5472.CAN-11-0898

Figure 2. Pemetrexed interacts with sorafenib in a dose-dependent fashion to increase autophagy and tumor cell killing that is suppressed by knock down of Beclin1.

Figure 2

Figure 2

Panel A. BT474, 4T1 and HuH7 cells as indicated are transfected with siRNAs (si-scramble, siSCR; siBeclin1; 20 nM) and with a plasmid to express LC3-GFP. Twenty four h after transfection cells were treated with vehicle (PBS) or pemetrexed (0.03–3.0 µM) and/or vehicle (DMSO) or sorafenib. Twelve h after drug exposure cells were examined under a fluorescent microscope (X40) at the indicated times after drug exposure and the mean number of vesicles in 40 random cells in triplicate calculated per experiment (n = 2 studies, +/− SEM; * p < 0.05 less than corresponding siSCR value). Panel B. HuH7 and H460 cells were treated as indicated with vehicle (PBS) or pemetrexed and/or vehicle (DMSO) or sorafenib. Viability was determined in triplicate 24h later by trypan blue exclusion (HuH7, H460) or annexin-PI staining, as noted in the panel (n = 2, +/− SEM; ¶ p < 0.05 greater than vehicle control value). Panel C. Parental MCF7 and fluvestrant resistant MCF7 cells (MCF7F) were treated as indicated with vehicle (PBS) or pemetrexed and/or vehicle (DMSO) or sorafenib. Viability was determined in triplicate 24h later by trypan blue exclusion (n = 2, +/− SEM; ¶ p < 0.05 greater than vehicle control value; ¶¶ p < 0.05 greater than corresponding value in parental MCF7 cells). Panel D. BT474 and HuH7 cells as indicated are transfected with siRNAs (si-scramble, siSCR; siBeclin1; 20 nM). Twenty four h after transfection in triplicate cells were treated with vehicle (PBS) or pemetrexed (0.03–3.0 µM) and/or vehicle (DMSO) or sorafenib. Viability was determined in triplicate 24h later by trypan blue exclusion (n = 2, +/− SEM; * p < 0.05 less than corresponding siSCR value).