Posttranslational Modification Defects in Fibroblast Growth Factor Receptor 1 as a Reason for Normosmic Isolated Hypogonadotropic Hypogonadism

Figure 3

Analysis of subcellular localization and deglycosylation in FGFR1 groups. (a) Subcellular localization of FGFR1 in HEK293 cells. HEK293 cells were transfected with empty vector (EV), FGFR1 (WT), or mutant FGFR1 (p. E670K), and protein localization was observed by fluorescence microscopy. FGFR1 was detected using an anti-FGFR1 antibody followed by secondary antibodies conjugated with Alexa Fluor 555 (red). Nuclei were visualized by DAPI. Original magnification: 600x. (b) Deglycosylation in FGFR1 groups with Endo H and PNGase F. Overall expression levels of FGFR1 in distinct groups were judged from the PNGase F treatments and were normalized to their GAPDH levels, respectively. Maturation analysis was determined from the Endo H-treated groups; the upper band represents the fully glycosylated mature form while the lower band stands for an immature or a core glycosylated form. Percentage of the mature band by density calculations was used to measure the maturation degrees with the groups. Both the results represent the ratio between mutant and WT. EV: empty vector; UT: untreated; E: Endo H-treated; P: PNGase F treated. Arrows point to the molecular weight.

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