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. 2011 Sep 20;20(1):46–53. doi: 10.1038/mt.2011.197

Figure 4.

Figure 4

Lenti-HF2AL infection results in tumor cell death by apoptosis in vitro. (a) Cytotoxicity of lenti-HF2AL in various tumor cell lines, including SMMC7721, BEL7402, HepG2, HCT116, MDA-MB-231, U937, SVT35, A549, and U251 and two normal cell lines, HFTF and HEK 293. Tumor cells or normal cells were infected with lenti-HF2AL at a MOI of 25 for 24 hours, and the media were then replaced with fresh media, and cell viability was determined by an MTS assay 48 hours later. Replicates were averaged, and the error bars represent the SEM of three independent experiments. (b) HCT116 cells infected with the indicated recombinant vectors (MOI = 25) were stained with Hoechst and examined under fluorescent microscopy. Arrows indicate typical apoptotic cells exhibiting nuclear condensation. (c) Caspase-8, -9, and -3 activation in HCT116 cells treated with conditional medium from lenti-HF2AL-infected HEK 293 cells at the indicated time point was detected by western blot analysis using corresponding antibodies. GAPDH was used as the loading control. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MOI, multiplicity of infection; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; PBS, phosphate-buffered saline.