Fig. 6

Circ_005077 acted as a scaffold to promote the binding of CyPA and p47phox and activated NADPH oxidase. (A) The binding of CYPA to p47phox was determined by Co-IP and Western blotting in H9C2 cells with or without rno_circ_005077 overexpression. (B) Flag-CyPA and HA-p47phox were co-transfected and expressed in H9c2 cells with or without overexpression of circ_005077; Co-IP and Western blotting determined the binding of Flag-CyPA to HA-p47phox. (C, D) Protein levels of p47phox in H9c2 cells with inhibition or overexpression of circ_005077 were shown. (E,F) The ELISA assays showed the NOX activity of H9C2 cells with overexpression or inhibition of circ_005077, and inhibition of CyPA or overexpression of p47phox can rescue this effect under PA.(G,H) NADPH and the NADPH/NADP⁺ ratio were measured in H9c2 cells with overexpression of circ_005077, and inhibition of CyPA can reverse this impact under PA.(I,J) The levels of NADPH and the NADPH/NADP⁺ ratio were measured in H9c2 cells with inhibition of circ_005077, and p47phox overexpression can reverse this impact under PA.Data were shown as mean ± SD of at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001. NS, no significance