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. 2015 May 22;6(25):21283–21300. doi: 10.18632/oncotarget.4238

Figure 4. Transient inhibition of FUCA-induced autophagic cell death in MDA-MB-231 cells.

Figure 4

A. FUCA1 protein expression in MDA-MB-231 cells was transiently inhibited using siRNA. A DNA fragmentation assay was performed. MDA-MB-231 cells treated with Etoposide (EP, 50 μM) and griseofulvin (GF, 60 μM) were used as positive controls [48]. B. Cellular morphology was observed after transient transfection of the FUCA-Si plasmid. The cells appeared to lose cell-cell contact, and cytoplasmic vacuoles became apparent. C. Transient FUCA-knock-down cells were subjected to acridine orange staining to detect the formation of AVOs [22]. The autophagic cell death values were calculated as the percentage of acridine orange-positive cells relative to the total number of cells in each random field, and the data are presented as the mean ± SD from 3 independent experiments (*P = 0.001). D. MDA-MB-231 cells were transiently transfected with FUCA-Si plasmids for 24 h. Immunoblotting analyses for LC3, Beclin 1, and p62/SQSTM1 expression were performed as described in the Materials and Methods section.