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. 2016 Oct 16;7(50):82273–82288. doi: 10.18632/oncotarget.12701

Figure 3. c-Myc expression is CE-dependent.

Figure 3

(A) IMEC/vec and IMEC/c-Myc were transfected with CE siRNA or non-targeting control for 72hrs. CE transcript expression normalised to GAPDH was determined by RT-qPCR. Error bars represent standard error of the mean, n=3 (B) IMEC/vec and IMEC/c-Myc transfected with CE siRNA (+) or non-targeting control (−) for 72hrs. Protein was analysed by Western blotting. (C) Densitometry was performed to quantify the decrease in c-Myc Western blot signal (normalised to actin) following CE depletion, relative to control transfection. Error bars represent standard error of the mean, n=6. (D) IMEC/vec and IMEC/c-Myc were stably transduced with GFP or a CE-GFP construct containing wobble codons to confer resistance to CE siRNA. Cells were transfected with CE siRNA (+) or control (−) and protein analysed by Western blot. (E) HeLa cells were transfected with two independent CE siRNAs (#1 or #2) or non-targeting control (−) for 72hrs. Protein analysed by Western blotting. (F) IMEC/vec, IMEC/c-Myc and HeLa cells were transfected with CE siRNA or non-targeting control. Endogenous c-Myc and Flag-c-Myc transcript level normalised to GAPDH was determined by RT-qPCR. Error bars represent standard error of the mean, n≥6. Significance relative to control siRNA calculated by Student's T-test, *** = p≤0.001; ** = p≤0.01; * = p≤0.05.