Fig. 3: NGC blocks COL6A3 cleavage without affecting MMP catalytic activity. | Experimental & Molecular Medicine

Fig. 3: NGC blocks COL6A3 cleavage without affecting MMP catalytic activity.

From: Targeting COL6A3-C5 with nigericin suppresses endotrophin formation and enhances insulin sensitivity in obesity

Fig. 3: NGC blocks COL6A3 cleavage without affecting MMP catalytic activity.

a–d Luciferase activities of the ETP-23AA cleavage biosensor were determined using multiple catalytic MMPs (caMMP1 (a), caMMP2 (b), caMMP9 (c) and caMMP16 (d)). Statistical significance was evaluated using one-way ANOVA. *P < 0.05 (n = 3 per group). e, f Activity of MMPs was measured in conditioned medium using gelatin zymography. Conditioned media of HEK293T cells, transiently transfected with caMMP2 (e), or caMMP9 vector (f), were incubated with the indicated NGC for 16 h and analyzed using zymography. g Validation of NGC specificity on ETP cleavage activity with C2C5–FLAG constructs containing point mutations at cleavage sites (L/M). ETP, cleaved from C2C5 by endogenous MMPs, was detected using immunoblotting with anti-FLAG antibodies and suppressed by NGC. The L/M mutant failed to release ETP regardless of NGC treatment.

Back to article page