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. 2014 Feb 17;124(3):1098–1113. doi: 10.1172/JCI69778

Figure 8. Calpain-induced talin1 cleavage in injured podocytes.

Figure 8

(A) Glomerular lysate isolated following protamine sulfate (PS) infusion into mice demonstrates talin1 cleavage (mouse anti-talin C terminus, clone TD77; Millipore). (B) Injection of NTS in mice demonstrates rabbit IgG deposition in the GBM. Scale bar: 10 μm. Alexa Fluor goat anti rabbit denoted by gαr. Alexa Fluor goat anti-mouse, denoted by gαm, demonstrates no signal. (C) Talin1 cleavage is observed in glomeruli following injection of NTS in mice. (D) Glomerular calpain activity is increased following protamine sulfate infusion, n = 7 experiments. (E) Glomeruli isolated from mice injected with NTS exhibit increased calpain activity, n = 5 experiments. (F) Increased urinary calpain activity is observed following NTS stimulation. n = 10 experiments. (G) Supernatant obtained from primary podocytes treated with protamine sulfate exhibit increased calpain activity. n = 3 experiments. (H) Podocytes stimulated with protamine sulfate (200 μg/ml) demonstrate increased talin1 and spectrin cleavage. (I) Isolated primary proximal tubules show no increase in talin1 cleavage following NTS injection in mice. (J) Isolated primary proximal tubules show no increase in calpain activity following NTS injection in mice. n = 4. *P < 0.05; **P < 0.01.