The Role of CD147 in Pathological Cardiac Hypertrophy Is Regulated by Glycosylation

Figure 1

CD147 expression in mouse hearts. Mice overexpressing wild-type CD147 or glycosylation defective CD147 were generated by injecting adeno-associated virus-9 vectors containing the individual constructs into the myocardia. (a) Representative western blot analysis of CD147 expression in mouse hearts treated in the presence or the absence of peptide-N-glycosidase F (PNGase F). Three bands of approximately 32 kDa, 38 kDa, and 50 kDa were detected, which corresponded to nonglycosylated (core protein, NG), low-glycosylated (LG), and high-glycosylated (HG) CD147, respectively. (b) Western blot analysis of CD147 expression in mouse hearts after sham or transverse aortic constriction surgery (/group). (c) Mutagenesis of CD147 glycosylation sites. Three conserved asparagine (Asn) glycosylation sites (N44, N154, and N190) in mouse CD147 were converted to glutamine (Gln) to generate glycosylation-defective CD147 protein. (d) Western blot analysis of overexpression of CD147-WT-3XFlag and CD147-mutant-3XFlag in mouse hearts 14 days after injection with AAV-9 vector alone (vehicle), AAV-9 with wild-type CD147 (CD147-WT), or AAV-9 with glycosylation-defective CD147 (CD147-Mut). It is noteworthy that the bands for glycosylated CD147 (CD147-WT) and mutant nonglycosylated CD147 (CD147-Mut) shifted up in the immunoblot because of the addition of the 3XFlag protein. vs. the sham group; ns: not significant.

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