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

Figure 4

Functional analysis of FGFR1 in the mutant and WT groups in vitro. (a) Gene expression analysis of FGFR1 in the mutant and WT groups by qPCR. HEK293 cells were transiently transfected with WT, mutant FGFR1 plasmid, or empty vector for RNA extraction, using RT-PCR and real-time quantitative PCR to detect the FGFR1 mRNA expression. (b–d) Gene and cell surface expression analysis of FGFR1 and its downstream signaling. (b) Using RT-PCR and qPCR for the FOS gene expression analysis in the FGF8-induced mutant and WT groups. (c) The phosphorylation levels of WT and mutant FGFR1 and the affected signal pathways tested by western blotting in groups (A). Quantitative analyses of FGFR1 phosphorylation levels (Y653) (B), the total FGFR1 relative level (C), FGF8-induced ERK1/2 (D), and Akt (E) phosphorylation levels are shown with a bar chart. (d) Analysis on the FGFR1-affected JAK/STAT3 pathways by western blotting in groups (A). Quantitative analysis of the STAT3 phosphorylation levels (B). Shown is the of three biological replicates ( by Student’s -test).

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