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. 2014 Nov;5(11-12):407–419. doi: 10.18632/genesandcancer.44

Figure 1. DEPTOR knockdown induces Apoptosis, G1/S cell cycle arrest & senescence in MM cells.

Figure 1

A). Cell lines acutely infected with control (scramble) or two separate DEPTOR shRNAs followed by Western blot. Phospho-P70/total P70 ratio shown under the gel was determined by densitometry and is mean of 3 separate experiments; B) % apoptosis 3 or 4 days after lentivirus infection with control (scr) or shRNA to DEPTOR (sh1, sh2). C) 8226 transfected with dox-inducible control shRNA (Luc or GFP) or shRNA targeting 2 sequences of DEPTOR (DEPTOR-1/DEPTOR-2); Dox added to induce shRNAs and immunoblot 72 hrs later for DEPTOR expression or PARP cleavage (arrow); D) Immunoblot assay performed in 8226 cells at 1, 2 or 3 days after adding dox; E) MTT assay over time after adding dox to induce shRNA; F) Apoptosis assay at 72 hrs (flow analysis of activated caspase 3) in control (shLuc) or DEPTOR knockdown cells (shDEP) +/− dox. ZVAD added at 50 or 100 uM. G) G1 or S distribution in transfected 8226+/− dox. H) Senescence assay (SA-beta gal staining (arrows)) in DEPTOR knockdown cells after adding dox; I) Quantification of senescence. All data in figs 1B, 1E, 1F, 1G and 1I represent means+/−SD, n=3).