Fig. 2.

Top: Enzyme-linked immunoabsorbent (ELISA) evaluation of caspase-3, caspase-8, and caspase-9 activities on tubular cells cultured for 48 h with 5% CONV or PMX plasma. PMX T0 and CONV T0 and T72 plasmas induced a significant increase of all caspase activities (*P < 0.05 vs. healthy plasma). A significant decrease of all caspase activities was found with PMX T72 plasma compared to PMX T0 (^† P < 0.05 PMX T72 vs. PMX T0); however, caspase-3 and caspase-9 activities remained significantly higher than healthy plasma (*P < 0.05 PMX T72 vs. healthy plasma). Middle: Representative images of FACS and immunofluorescence (insets) analysis of Fas (CD95) expression on tubular cell surface after exposure to CONV or PMX plasma. PMX T0 and CONV T0 and T72 plasmas all induced a marked upregulation of Fas, which was significantly reduced in presence of PMX T72 plasma. ×400 magnification. Bottom: Representative western blot analysis of the mitochondrial proteins Bax and Bcl2 in tubular cells exposed to CONV or PMX plasma, and related densitometric analysis expressed as Bax/Bcl2 ratio. PMX T0, CONV T0, and CONV T72 plasmas induced a marked upregulation of the Bax/Bcl2 ratio that was reduced in the presence of PMX T72 plasma. (Lane 1 Vehicle; Lane 2 Healthy; Lane 3 PMX T0; Lane 4 PMX T72; Lane 5 CONV T0; Lane 6 CONV T72). Beta-actin was used as reference for protein loading