Hepatocyte cytoskeleton during ischemia and reperfusion - influence of ANP-mediated p38 MAPK activation

Figure 2 Influence of ANP preconditioning and ischemia on total actin and F-actin content. Livers were perfused for 30 min in the absence or presence of 200 nmol/L ANP, which was added 20 min prior to ischemia. Afterwards, livers were kept under ischemic conditions for 24 h (4 °C) in UW solution. At the indicated times, livers were snap-frozen, homogenized, and analyzed by Western blot using an anti-actin antibody. For differentiation between F- and G-actin (cytoskeleton and cytosol, respectively), liver homogenates were fractionated before Western blot analysis according to “Materials and methods” (A and B). One representative Western blot is imaged for each experimental setting. (A) Western blots representing hepatic F- and G-actin content during IR in untreated (Co) and ANP-preconditioned livers after 30-min perfusion (30' perf.), 24-h ischemia (24-h isch.), and 45-min reperfusion (45' rep.). (B) Quantitative/densitometric analysis of F- and G-actin content during IR in untreated (Co) or ANP-preconditioned livers. Data are expressed as percent of total actin content (F-actin+G-actin=100%) as mean±SE of n = 4 experiments in each treatment group. ^bP<0.001 vs appropriate control. (C) Western blot showing total actin in untreated liver homogenates after 30-min perfusion, 24-h ischemia, and 45-min reperfusion. (D) Total hepatic actin content after perfusion (30') and ischemia (24-h) in ANP treated or untreated (Co) livers detected by Western blot analysis.