Figure 5
Effect of drug treatments on markers of autophagy and apoptosis. (A) LC3 immunofluorescence staining of KYSE450 cells treated with 5-FU (40 μ M) alone or in combination with HA14-1 (20 μ M) for 24 and 48 h. Untreated KYSE450 cells displayed a diffuse LC3 distribution (upper left panel), in contrast, following treatment, staining was bright and punctuate (all other panels). (B) The extent of active caspase 3 was analysed by fluorescence microscopy in (i) KYSE450 and (ii) OE21 cells treated with a range of concentrations of 5-FU (KYSE450 cells 20–40 μ M 5-FU; OE21 cells 20–30 μ M 5-FU) and HA14-1 (20 μ M) for 48 and 24 h, respectively. Cells exhibiting active caspase 3 were quantified by counting five fields of view per slide, with ∼30 cells per field ( × 40 magnification). Representative images of caspase 3 are shown on the right-hand panels.
