Fig. 2

Genetic and expression analysis of synonymous mutations c.414A > G using primary fibroblasts. (a) Schematic of the exon structures of human VHL gene with the positions of the mutation and two RT-PCR primers indicated. (b) Chromatogram tracks showing the DNA sequence near the mutation site for each of the five fibroblast cell lines established from the patient family. (c) RT-PCR bands amplified from the cDNA of E1E2E3 and E1E3 transcripts using mRNA extracted from fibroblast cells. GAPDH was used as an internal control. (d) Western blot showing the three pVHL isoforms in the five fibroblast cell lines. RCC cell line 786O (VHL-null) and HEK293T (VHL-intact) were used as control for pVHL detection. α-Tubulin was used as loading control. VHL antibody is from Cell Signaling (cat#68547)