Fig. 2
From: GEN1 as a risk factor for human congenital anomalies of the kidney and urinary tract
Nuclease activity of GEN1 variant proteins at HJs. As a HJ enzymatic lyase, GEN1 protein’s dissociation ability was partly manifested by the degradation of the equal HJ (substrate) by different concentrations of purified proteins. In this experiment, seven gradient concentrations were applied (0, 8, 16, 32, 64, 128 and 256nM). GEN1 (p.R401X,508) showed the most significant reduction in enzymatic hydrolytic capacity. The other five mutant proteins [GEN1 (p.T105R), GEN1 (p.D149N), GEN1 (p.H369L), GEN1 (p.I577T), and GEN1 (p. T105R + p.I577T)] impaired the enzymatic hydrolysis of HJ. The impaired enzymatic hydrolytic capacity in the control group variants was less significant than that in CAKUT group. (A) Grayscale statistics for the above grayscale map, reflecting the comparison of the ability of wild-type and mutant GEN1 protein to degrade HJ. All results of all mutated proteins were evaluated from the baseline results of wild-type proteins. (B) Grayscale statistical chart. The results are reported as mean ± SD for three individual experiments. * sites in the non-CAKUT group. Abbreviations: EMSA, electrophoretic mobility shift assay; HJ, Holliday junction; WT, wild type