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Expression of CNPY2 in mouse tissues: quantification and localization.

Hatta K, Guo J, Ludke A, Dhingra S, Singh K, Huang ML, Weisel RD, Li RK - PLoS ONE (2014)

Bottom Line: CNPY2 was also detectable in mouse blood and human and mouse uteri.These data demonstrate CNPY2 is widely distributed in tissues and suggest the protein has biological functions that have yet to be identified.Using these new observations we discuss possible functions of the protein.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiovascular Surgery, Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada; Institute of Medical Sciences, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT
Canopy FGF signaling regulator 2 (CNPY2) is a FGF21-modulated protein containing a saposin B-type domain. In vitro studies have shown CNPY2 is able to enhance neurite outgrowth in neurons and stabilize the expression of low density lipoprotein receptor in macrophages and hepatocytes. However, no in vivo data are available on the normal expression of CNPY2 and information is lacking on which cell types express this protein in tissues. To address this, the present study examined CNPY2 expression at the mRNA and protein levels. Quantitative PCR and ELISA examination of mouse tissues showed that CNPY2 varies between organs, with the highest expression in the heart, lung and liver. Immunohistochemistry detected CNPY2 in a variety of cell types including skeletal, cardiac and smooth muscle myocytes, endothelial cells and epithelial cells. CNPY2 was also detectable in mouse blood and human and mouse uteri. These data demonstrate CNPY2 is widely distributed in tissues and suggest the protein has biological functions that have yet to be identified. Using these new observations we discuss possible functions of the protein.

No MeSH data available.


Western blotting and ELISA of CNPY2.A. Recombinant His-CNPY2 protein was purified from E. coli to generate the rabbit polyclonal antibody. The purified protein is shown on a Coomassie-stained gel. B. In a Western blot, the antibody was tested for detection of CNPY2 fusion proteins (CNPY2-GFP, ∼47 kDa and CNPY2-V5-His, ∼23 kDa, respectively) in the cell lysates of HEK293 cells after transient transfection. C. The anti-CNPY2 antibody was used to test the expression of native CNPY2 in mouse heart tissue. Anti-GAPDH served as a positive control. D. ELISA was performed to quantify CNPY2 protein abundance in tissues.
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pone-0111370-g002: Western blotting and ELISA of CNPY2.A. Recombinant His-CNPY2 protein was purified from E. coli to generate the rabbit polyclonal antibody. The purified protein is shown on a Coomassie-stained gel. B. In a Western blot, the antibody was tested for detection of CNPY2 fusion proteins (CNPY2-GFP, ∼47 kDa and CNPY2-V5-His, ∼23 kDa, respectively) in the cell lysates of HEK293 cells after transient transfection. C. The anti-CNPY2 antibody was used to test the expression of native CNPY2 in mouse heart tissue. Anti-GAPDH served as a positive control. D. ELISA was performed to quantify CNPY2 protein abundance in tissues.

Mentions: Recombinant CNPY2 (Figure 2A) was purified and used to generate antibody. The antibody was used to visualize CNPY2-GFP and CNPY2-V5-His on a Western blot and the predicted molecular weight bands for the respective fusion proteins were detected (Figure 2B). Native mouse CNPY2 was detected in mouse heart tissue by Western blotting (Figure 2C) and was measured in all mouse tissues examined by ELISA (Figure 2D). However, higher CNPY2 expression levels were observed in lung, heart and liver. CNPY2 was also detectable in mouse blood plasma by ELISA at a concentration of 120.0±1.4 pg/mL (n = 5).


Expression of CNPY2 in mouse tissues: quantification and localization.

Hatta K, Guo J, Ludke A, Dhingra S, Singh K, Huang ML, Weisel RD, Li RK - PLoS ONE (2014)

Western blotting and ELISA of CNPY2.A. Recombinant His-CNPY2 protein was purified from E. coli to generate the rabbit polyclonal antibody. The purified protein is shown on a Coomassie-stained gel. B. In a Western blot, the antibody was tested for detection of CNPY2 fusion proteins (CNPY2-GFP, ∼47 kDa and CNPY2-V5-His, ∼23 kDa, respectively) in the cell lysates of HEK293 cells after transient transfection. C. The anti-CNPY2 antibody was used to test the expression of native CNPY2 in mouse heart tissue. Anti-GAPDH served as a positive control. D. ELISA was performed to quantify CNPY2 protein abundance in tissues.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4230931&req=5

pone-0111370-g002: Western blotting and ELISA of CNPY2.A. Recombinant His-CNPY2 protein was purified from E. coli to generate the rabbit polyclonal antibody. The purified protein is shown on a Coomassie-stained gel. B. In a Western blot, the antibody was tested for detection of CNPY2 fusion proteins (CNPY2-GFP, ∼47 kDa and CNPY2-V5-His, ∼23 kDa, respectively) in the cell lysates of HEK293 cells after transient transfection. C. The anti-CNPY2 antibody was used to test the expression of native CNPY2 in mouse heart tissue. Anti-GAPDH served as a positive control. D. ELISA was performed to quantify CNPY2 protein abundance in tissues.
Mentions: Recombinant CNPY2 (Figure 2A) was purified and used to generate antibody. The antibody was used to visualize CNPY2-GFP and CNPY2-V5-His on a Western blot and the predicted molecular weight bands for the respective fusion proteins were detected (Figure 2B). Native mouse CNPY2 was detected in mouse heart tissue by Western blotting (Figure 2C) and was measured in all mouse tissues examined by ELISA (Figure 2D). However, higher CNPY2 expression levels were observed in lung, heart and liver. CNPY2 was also detectable in mouse blood plasma by ELISA at a concentration of 120.0±1.4 pg/mL (n = 5).

Bottom Line: CNPY2 was also detectable in mouse blood and human and mouse uteri.These data demonstrate CNPY2 is widely distributed in tissues and suggest the protein has biological functions that have yet to be identified.Using these new observations we discuss possible functions of the protein.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiovascular Surgery, Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada; Institute of Medical Sciences, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT
Canopy FGF signaling regulator 2 (CNPY2) is a FGF21-modulated protein containing a saposin B-type domain. In vitro studies have shown CNPY2 is able to enhance neurite outgrowth in neurons and stabilize the expression of low density lipoprotein receptor in macrophages and hepatocytes. However, no in vivo data are available on the normal expression of CNPY2 and information is lacking on which cell types express this protein in tissues. To address this, the present study examined CNPY2 expression at the mRNA and protein levels. Quantitative PCR and ELISA examination of mouse tissues showed that CNPY2 varies between organs, with the highest expression in the heart, lung and liver. Immunohistochemistry detected CNPY2 in a variety of cell types including skeletal, cardiac and smooth muscle myocytes, endothelial cells and epithelial cells. CNPY2 was also detectable in mouse blood and human and mouse uteri. These data demonstrate CNPY2 is widely distributed in tissues and suggest the protein has biological functions that have yet to be identified. Using these new observations we discuss possible functions of the protein.

No MeSH data available.