Skip to main content
. 2016 Feb 1;2:16004. doi: 10.1038/cddiscovery.2016.4

Figure 5.

Figure 5

Bcl-xL and cFLIP expression block TRAIL-induced apoptosis in NGF-sensitized TrkA SH-SY5Y cells and cFLIP is responsible for the TRAIL-resistant SH-SY5Y phenotype. (a) Western blot demonstrating transient Bcl-xL overexpression in TrkA SH-SY5Y cells transiently transfected with a mammalian pcDNA3.1 Bcl-xL expression vector but not with empty pcDNA3.1 vector. (b) Histogram demonstrating significant (*) inhibition of TRAIL-induced apoptosis in NGF-treated Bcl-xL-transduced but not empty vector-transfected TrkA SH-SY5Y cells in response to NGF (100 ng/ml) plus TRAIL (200 ng/ml). (c) Representative western blots demonstrating cFLIP-specific siRNA but not control siRNA knockdown of cFLIP but not α-tubulin in TrkA SH-SY5Y cells. (d) Histogram demonstrating a significant (*) increase in apoptosis of TrkA SH-SY5Y cells exhibiting siRNA cFLIP knockdown (cFLIP siRNA) but not in cells transiently transfected with control siRNA duplexes (Con SiRNA) in the presence but not absence of TRAIL and in the absence of NGF. (e) Representative western blot demonstrating transient overexpression of FLAG-tagged cFLIP in TrkA SH-SY5Y cells but not in cells transfected with empty pcDNA expression vector. (f) Histogram demonstrating a significant (*) reduction in TrkA SH-SY5Y apoptosis induced by NGF (100 ng/ml) plus TRAIL (200 ng/ml) in TrkA SH-SY5Y cells that transiently overexpress exogenous FLAG-tagged cFLIP compared with empty expression vector-transfected counterparts. Histogram results are displayed as the mean percentage (%) (±S.D.) alive (white) or dead (black) cells in three experiments, each performed in duplicate (*statistical significance).