Journal: International Journal of Molecular Sciences
Article Title: A Pre-Clinical Study on the Use of the Proprotein Convertase Subtilisin/Kexin Type 9 Inhibitor PEP 2-8 to Mitigate Ischemic Injury in a Rat Marginal Donor Model
doi: 10.3390/ijms26188937
Figure Lengend Snippet: PEP 2-8 effect on cell proliferation and apoptosis in control kidneys (CTRL, n = 15) and PEP 2-8 treated kidneys (PEP 2-8, n = 15). ( A ) Top: Box plots represent tubular proliferation index (TPI). TPI was defined as the ratio between the nuclei expressing PCNA and the total nuclei in each tubule, in ten non-consecutive fields from each immunostained kidney (×40 magnification). Data are expressed as median and 2.5–97.5 percentile. Wilcoxon test was performed to compare data from the two study groups. (* p < 0.05) Bottom: Representative sections of PCNA immunostaining (×40 magnification). ( B ) Top: Apoptosis frequency in the two study groups was assessed by counting a total of 1000 tubular cell nuclei per kidney. Results are presented as the percentage of kidneys with cleaved caspase-3 positive (pink) or negative (white) staining. Data from the two groups were compared using Fisher’s exact test. (**** p < 0.0001) Bottom: Representative sections of cleaved caspase 3 immunostaining (40× magnification).
Article Snippet: Following three washes in 150 mM PBS, antigen retrieval was performed by microwaving the sections in citrate buffer pH 6 and then incubated overnight at 4 °C with monoclonal mouse anti-proliferating cell nuclear antigen (PCNA) antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or N-Tyrosine (N-Tyr) antibody (1:100 Santa Cruz Biotechnology, Inc., Dallas, TX, USA).
Techniques: Control, Expressing, Immunostaining, Staining