Figure 1.
Immunohistochemistry in Cmah-dKO mouse-derived tissues (lung, kidney, and heart) for detection of CMAH and Neu5Gc.
WT mice expressed both Cmah and Neu5Gc epitopes. Cmah-dKO mice were completely deficient for both CMAH and Neu5Gc epitopes in the lung, kidney, and heart tissues. Bar: 100 µm. LU; lung, KI; kidney, HE; heart.
Figure 2.
Sialyltransferase gene expression levels in WT and Cmah-dKO mice.
A. Comparison of sialyltransferase gene expression in the liver of WT and Cmah-dKO mice by RT-qPCR. B. RT-qPCR analysis in liver of WT and Cmah-dKO mice. All RT-qPCR were conducted in triplicate and normalized with mouse Gadph gene expression. The data were presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001.
Figure 3.
Gene expression profile in the lung, kidney, and heart of Cmah null mice.
A. Venn diagram showing differential expression of genes in the lung, kidney, and heart of Cmah null mice. Numbers in red and blue Venn diagram present up- and down- regulated genes, respectively. LU, lung; KI, kidney; HE, heart. B. The differentially up- or down-regulated genes were classified according to biological process in the lung, kidney, and heart of Cmah-dKO mice. LU, lung; KI, kidney; HE, heart. X axis of bar graph indicates the gene numbers.
Table 1.
Top 6 biological processes overrepresented in the lung, kidney, and heart of Cmah null mice.
Table 2.
KEGG pathways significantly associated with genes altered in the lung, kidney, and heart of Cmah null mice.
Figure 4.
Networks predicted by Ingenuity Pathway Analysis in the Cmah-dKO mice.
Upper: The common network identified was lipid metabolism and small-molecule biochemistry in the lung (LU), kidney (KI), and heart (HE), respectively. The network is displayed graphically as nodes (genes). The node color intensity indicates the expression of genes, with red representing up-regulation and green representing down-regulation. Solid lines and dotted lines indicate direct relationship and indirect relationships, respectively. Bottom: The table includes the molecules, score, and focus molecules in identified common networks (lipid metabolism and small molecule biochemistry) from each tissue by IPA (score 30, 13 focus molecules in LU; score 48, 18 focus molecules in KI; score 34, 14 focus molecules in HE). Red and blue denote up-regulated and down-regulated genes among the differentially expressed genes.
Table 3.
Disease and disorders predicted by Ingenuity Pathway Analysis in lung, kidney, and heart of Cmah null mice.
Figure 5.
Schematic representation of biological networks was constructed by CMAH function.
A. Validation of microarray gene expression by RT-qPCR. Gene expression levels of selected genes were determined by RT-qPCR using WT and Cmah-dKO derived lung, kidney, and heart tissues. Measurements were performed in triplicate, after which the calculated mean expression level was corrected for Gadph gene expression levels. Error bars indicate standard deviations. B. Functional correlation and interaction of both the signaling proteins and Cmah gene were reconstituted into a network model using the Pathway Studio software (Ariadne Genomics Inc.; Rockville, MD, USA). All genes are shown by their gene symbols. Direct and indirect regulation is indicated by colored lines and dotted gray lines, respectively.