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. 2013 Dec 3;8(12):e81928. doi: 10.1371/journal.pone.0081928

Figure 5. Effect of PARP inhibitor on the PI-3-kinase – Akt pathway and Akt level in transplanted and control kidney samples.

Figure 5

(A) Effects of PARP inhibitor on the Akt1 protein, and phosphorylation of Akt1 and GSK-3 β in control and transplanted kidneys determined using immunoblotting with protein and phospho-specific primary antibodies. Actin was used as loading control. Representative blots of at least three parallel experiments are presented. B. The bar diagrams represent pixel volumes of phosphorylated Akt (serine 473) in kidney samples. The bands were normalized to the appropriate actin band. *p<0.01 transplanted PARP inhibitor treated samples compared to other samples. Difference between control samples (independently of PARP treatment) and transplanted untreated kidney samples were not significant. The bar diagrams represent pixel volumes of GSK-3β (serine 9) phosphorylation bands. The bands were normalized to the appropriate actin band. *p<0.001 transplanted PARP inhibitor treated samples compared to control PARP inhibitor treated, or untreated samples. **p<0.05 untreated transplanted kidney samples compared to control samples (independently of PARP treatment. *** p<0.01 transplanted PARP inhibitor treated samples compared untreated transplanted samples. The bar diagrams represent pixel volumes of Akt1 protein bands. The bands were normalized to the appropriate actin band. *p<0.001 transplanted kidney samples (independently of PARP inhibitor treatment) compared to control kidney samples (independently of PARP inhibitor treatment). (C) Effects of PARP inhibitor on nuclear NF-kappaB and p-NF-kappaB leveles in control and transplanted kidneys determined by immunoblotting with protein and phospho-specific primary antibodies. Actin was used as loading control. Representative blots of at least three parallel experiments are presented. (D) The bar diagrams represent pixel volumes of nuclear NF-kappaB protein bands. The bands were normalized to the appropriate actin band. *p<0.01 transplanted PARP inhibitor treated, or untreated, samples compared to control PARP inhibitor treated, or untreated control samples. **p<0.05 untreated transplanted kidney samples compared to PARP inhibitor treated transplanted kidney samples. The bar diagrams represent pixel volumes of nuclear p-NF-kappaB bands. The bands were normalized to the appropriate actin band. *p<0.001 transplanted PARP inhibitor treated, or untreated, samples compared to control PARP inhibitor treated, or untreated control samples. **p<0.05 untreated transplanted kidney samples compared to PARP inhibitor treated transplanted kidney samples. The vertical axes represent pixel volume means±SEM of the scanned bands of the immunoblots in arbitrary units.