Clinical and Functional Characteristics of a Novel KLF11 Cys354Phe Variant Involved in Maturity-Onset Diabetes of the Young

Figure 3

Functional analysis of the KLF11 Cys354Phe variant. (a) Protein expression of KLF11 (WT and Cys354Phe). Cell lysates of KLF11-expressing cells were used. Protein production was tested via western blotting. (b) Histogram of the KLF11 protein expression level analysis. (c) Luciferase assays of HEK 293 cells transfected with each KLF11 expression vector (WT and Cys354Phe). denotes () for the KLF11 WT plasmid compared to the empty plasmid; n.s. denotes () for KFL11/C354F compared to the empty plasmid; # denotes () for KFL11/C354F compared to KLF11 WT. (d–f) INS1 cells were transfected with the KLF11-WT or KLF11-Cys354Phe plasmids for 24 (d) or 48 h (e). (d) qRT-PCR was conducted to determine the Ins1 mRNA levels after stimulation with 2.8 mmol/L glucose (low) or 20 mmol/L glucose (high) for 24 h. (e) Insulin secretion levels in INS1 cells were analyzed via glucose-stimulated insulin secretion assay using ELISA after stimulation with 2.8 mmol/L glucose (low) or 20 mmol/L glucose (high) for 2 h. Note: denotes the number of experiments. denotes () for KLF11 WT compared to the empty plasmid; n.s. denotes () for KFL11/C354F compared to the empty plasmid; ## denotes () for KFL11/C354F compared to KLF11 WT.

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