Fig. 5: Dysfunction of genomic stability pathways by aberrant (P)RR expression in HPDE-1/E6E7 cells. | Communications Biology

Fig. 5: Dysfunction of genomic stability pathways by aberrant (P)RR expression in HPDE-1/E6E7 cells.

From: Aberrant (pro)renin receptor expression induces genomic instability in pancreatic ductal adenocarcinoma through upregulation of SMARCA5/SNF2H

Fig. 5

a Diseases expected from molecules downregulated by aberrant (P)RR expression. b Ingenuity Pathway Analysis (IPA) for molecular functions of the downregulated molecules under (P)RR overexpression. c Canonical pathways downregulated by (P)RR overexpression. d Canonical pathways identified with high IPA statistical confidence. Underline indicates molecules confirmed by western blot. e Constructs of deletion mutants in human (P)RR and confirmation of gene transfection in the vectors with each of Mock (M), FL(P)RR (FL), NTF of (P)RR (⊿C) and CTF of (P)RR (⊿N). Endogenous (P)RR was used as a loading control. f Expression of molecules involved in genomic stability pathways in cells with the deletion of each domain of (P)RR. Consistent results are obtained in three independent experiments. g Activation of Wnt components. Consistent results are obtained in three independent experiments. h Cell proliferative ability (mean ± SEM, N = 3 for each, *P < 0.05 vs. FL(P)RR, N.S., not significant). i DNA fibre assay. Cells were incubated sequentially with 5-chlorodeoxyuridine (CldU) and 5-iododeoxyuridine (IdU) for 20 min each. Upper: Representative images of DNA replication fork in cells (×800). Lower left: Quantitative evaluation of replication fork rates in cells (mean ± SEM, N = 100 for each, ***P < 0.0001 vs. Mock in CldU, N.S., not significant, †††P < 0.0001 vs. Mock in IdU). Lower right: Distribution of replication fork rate with CldU. j Single-cell gel electrophoresis in cells with the deletion of each domain of (P)RR at three-passage. Left: Representative images of DNA tail in cells (×100). Right: Quantification of the cells with DNA tails (N = 100 for each, ***P < 0.001 vs. Mock, N.S., not significant). The horizontal line inside the box plot is the median and the vertical lines protruding the box extend to the minimum and the maximum values, respectively. The vertical width of the central box shows the inter-quartile deviation. k Telomere length evaluated by Flow-FISH analyses (mean ± SEM, N = 3 for each, *P < 0.05).

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