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. 2016 Nov 4;7(49):80820–80841. doi: 10.18632/oncotarget.13098

Figure 4. Effects of TRAIL, GW441756, GSK2656157 and MG132 upon the expression of components of TRAIL-induced apoptosis.

Figure 4

(A) Western blots demonstrating the effect of TRAIL (200 ng/ml 0–12 hours) upon cFLIP, cBID, Caspase-8, Caspase-3 and Caspase-9 levels in whole cell extracts from NT SH-SY5Y (NT), pcDNA SH-SY5Y (pcDNA) and TrkAIII SH-SY5Y cells (TrkAIII) and Bcl-2, Bcl-xL, Mcl-1, DcR-1, DcR-2, DR-5 and DR-4 protein levels in whole cell extracts from TrkAIII SH-SY5Y cells (TrkAIII) (20 μg of whole cell extract loaded per sample). (B) Agarose gels demonstrating cFLIPS, cFLIPL, Bcl-2, Bcl-xL, Mcl-1 and GAPDH RT-PCR product levels generated from mRNAs purified from TrkAIII SH-SY5Y cells treated with 1 μM GW441756 (GW; 0, 1 and 6 hours). (C) Western blots demonstrating the inhibitory effect of the PERK inhibitor GSK2656157 (GSK; 0.1 and 1 μM for 24 hours) upon Mcl-1 but not cFLIP, Bcl-2, Bcl-xL or α-tubulin (α-Tub) expression and the inhibitory effect of the TrkA inhibitor GW441756 (GW: 0.1 and 1.0 μM for 24 hours) upon Bcl-2 and Bcl-xL but not Mcl-1 or α-tubulin expression and its stimulatory effect upon cFLIP expression in whole cell TrkAIII SH-SY5Y cell extracts (20 μg). (D) Histograms of the adjacent Western blots, displaying the results described in (C), expressed as percentage change in densitometric ratio to α-tubulin. (E) Western blots demonstrating the effects of MG132 (10 μM, 0–3 hours) upon cFLIP and α-tubulin protein levels in whole cell extracts from TrkAIII SH-SY5Y cells (20 μg).