Fig. 3

Functional analysis. A The NT5C3A gene expression analysis by quantitative PCR showed a very low expression level in patient MK compared with controls (healthy individuals). Error bars represent the standard deviation. B Western blot analysis of cytosolic pyrimidine 5′-nucleotidase expressed in erythrocyte (RK and MK—patients with HA, AK—asymptomatic father, EK—asymptomatic sister, and controls—healthy subjects). The volume corresponding to equal protein quantity of loaded samples is 25 µL. C Ponceau-S-stained nitrocellulose membrane as an alternative loading control. D Densitometric analysis of western blot assays for cytosolic pyrimidine 5′-nucleotidase present in the erythrocyte. The area of each peak (without background) for relative density calculation was used. Error bars represent range. Samples of all studied family members correspond to technical replicates (n = 2), while control samples correspond to biological replicates (n = 3)