Figure 1.
Relationship among and variability of samples.
Hierarchical clustering and multi-dimensional scaling analyses were performed. The MvA plots showing the small variation in A. WT/HT vs. KO/HT and B. WT/KID vs. KO/KID, and large variation in C. KO/HT vs. KO/KID and D. WT/HT vs. WT/KID.
Figure 2.
PAI-1 deficiency is associated with fibrosis in hearts but not in kidneys.
The levels of collagen accumulation in cardiac and renal tissues derived from wildtype and PAI-1 knockout mice were determined by Masson’s trichrome staining. Photographs were taken by an Olympus DP71 camera. A. 12-month old WT/HT; B. 12-month old PAI-1 KO/HT; C. 24-month old WT/HT; D. 24-month old PAI-1 KO/HT; E. 12-month old WT/KID, F. 12-month old PAI-1 KO/KID, G. 24-month old WT/KID, H. 24-month old PAI-1 KO/KID.
Figure 3.
Effect of PAI-1 deficiency on differential global gene expression profiling in hearts and kidneys.
Differentially expressed genes are presented by volcano plots with fold induction or repression and statistical significance in four different groups. A. Expression levels of genes in wildtype and PAI-1 knockout hearts. B. Expression levels of genes in wildtype and PAI-1 knockout kidneys. C. Expression levels of genes in PAI-1 knockout hearts and kidneys. D. Expression levels of genes in wildtype hearts and kidneys. E. Venn diagram showing the differentially expressed genes which are common in PAI-1 KO/HT vs. PAI-1 KO/KID group and WT/HT vs. WT/KID group (Blue); and unique in PAI-1 KO/HT vs. KO/KID group (Yellow) and WT/HT vs. WT/KID group (Red).
Table 1.
Differential gene expression in wildtype and PAI-1 knockout hearts.
Table 2.
Differential gene expression in wildtype and PAI-1 knockout kidneys.
Table 3.
Differential gene expression in PAI-1 knockout hearts and kidneys.
Table 4.
Differential gene expression in wildtype hearts and kidneys (Unique).
Figure 4.
Biological processes representing genes that were differentially expressed in wildtype and PAI-1 knockout heart and kidney groups.
Percentages of differentially expressed genes (upregulated or downregulated) under different biological processes are shown by Pie charts. A. WT/HT vs. PAI-1 KO/HT; B. WT/KID vs. PAI-1 KO/KID; C. PAI-1 KO/HT vs. PAI-1 KO/KID and D. PAI-1 KO/HT vs. PAI-1 KO/KID (unique); E. WT/HT vs. WT/KID; F. WT/HT vs. WT/KID (unique). Numbers of deregulated genes are shown in parentheses.
Table 5.
Biological Processes representing genes that were differentially expressed in knockout hearts compared to wildtype hearts (for level of expression of each gene see Table S1).
Table 6.
Biological Processes representing genes that were differentially expressed in knockout kidneys compared to wildtype kidneys (for level of expression of each gene see Table S2).
Table 7.
Biological Processes representing genes that were differentially expressed in knockout hearts compared to knockout kidneys (unique) (for level of expression of each gene see Table S5).
Table 8.
Biological Processes representing genes that were differentially expressed in wildtype hearts compared to wildtype kidneys (unique) (for level of expression of each gene see Table S6).
Figure 5.
Validation of expression levels of mRNAs in aged PAI-1 knockout and wildtype hearts and kidneys by qPCR analysis.
Total RNA from wildtype and PAI-1 knockout heart and kidney tissues were used for quantitation of A. Ankrd1; B. Pi16; C. Egr1: D. Dbp and E. Timp1 by qPCR analysis using gene specific primers. Data represents mean of triplicates ±SEM. P value of each sample was indicated in the Figure A–E.
Figure 6.
Levels of Ankrd1, Pi16, Timp4 and Timp1 in TGF-β-treated endothelial cells.
Mouse cardiac endothelial cells were treated with TGF-ß2 (10 ng/ml) for 7 days. At the end of incubation, control and EndMT-derived fibroblast-like cells were harvested and whole lysates were prepared. Equal amounts of proteins were subjected to Western blot using specific antibodies as indicated.
Figure 7.
Schematic diagram showing possible involvement of Ankrd1-Egr1 axis and a subset of profibrotic factors as igniters of cardiac fibrosis.
Along with other key profibrogenic factors, Ankrd1-Egr1 axis may play a pivotal role in initiation of cardiac-selective fibrosis; and kidney is protected from PAI-1 deficiency-induced fibrogenesis due to lack of these profibrotic regulators. Antifibrotic Klotho may also protect kidney from fibrogenesis.