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. 2017 Aug 24;8(49):85868–85882. doi: 10.18632/oncotarget.20696

Figure 4. IRES-mediated co-translation of Myc-ELAS1 and FLAG-p53 rescues the apoptosis-inducing function of ELAS1 in p53-deficient SAS cells after CPT treatment.

Figure 4

(A) A schematic presentation of cosmid DNA that allows IRES-mediated co-translation of Myc-ELAS1 and FLAG-p53 in SAS/Tet-On Adv cells, and a protocol for the transfection experiment. PCE, preparation of cell extract. (B) Wb was performed to examine expression of the indicated proteins in the absence (-) or presence (+) of cosmid DNA. Turquoise and green arrowheads indicate bands for FLAG-p53 and Myc-ELAS1 derived from expression of cosmid DNA, respectively. (C) FC analysis. SAS/Tet-On Myc-vector, Myc-ELAS1, SAS Adv + cosmid vector, or SAS Adv + cosmid construct (CMV_6Myc-ELAS1_IRES_FLAG-p53) cells were transfected in the absence (NT) or presence of treatment with 0.2 μM CPT for 48 h. (D) The bar graphs show the percentages of subG1 cells as determined by FC. Red and blue arrows correspond to the subG1 peak of FC data shown in Figure 4C. (E) Wb to examine expression of the indicated proteins. Turquoise arrowheads indicate the band for FLAG-p53.