Fig. 6

Identify the binding sites of circYap with PABP and eIF4G. The binding sites of circYap with PABP and eIF4G were predicted by NPDock. The interaction sites of protein–nucleic acid structures were calculated by HBPLUS and shown schematically in a diagram generated by the NUCPLOT. Docking model of circYap with PABP and eIF4G was shown as a circYap and PABP, b circYap and eIF4G (N-terminus), c circYap and eIF4G (C-terminus). RNA shows as pink and protein as green (PABP), indigo (eIF4G N-terminus), or blue (eIF4G C-terminus). d The secondary structure representation of circ-Yap highlighting the interactive sites with PABP (green), eIF4G (C-terminus) (blue) and eIF4G (N-terminus) (red). e The mutation was placed on the binding sites of circYap with Yap mRNA (Mut-1) or two proteins (Mut-2). Yap expression in cells transfected with plasmid containing vector, circYap, mutant binding sites (Mut-1 and Mut-2), or mutant non-essential region (Mut-3) were examined by western blotting. f Lysates from MDA-MB231 cells transfected with vector, circYap, Mut-1, Mut-2, or Mut-3 were incubated with circYap probe. The circYap (left) or Yap mRNA (right) pulled down by the probe was examined by real-time PCR. n = 4. **p < 0.01 compared to vector control. ^##p < 0.01 compared to circYap overexpressed cells. g Lysates from MDA-MB231 cells transfected with vector, circYap, Mut-1, Mut-2, or Mut-3 were incubated with Yap mRNA probe. Yap mRNA (left) or circYap (right) pulled down by the probe was examined by real-time PCR. n = 4. **p < 0.01 compared to vector control. ^##p < 0.01 compared to circYap overexpressed cells. h Lysates from MDA-MB231 cells transfected with vector, circYap, Mut-1, Mut-2, or Mut-3 were incubated with circYap probe. The pulled down PABP or eIF4G was examined by western blotting