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. 2019 Oct 28;20(21):5345. doi: 10.3390/ijms20215345

Figure 3.

Figure 3

Effects of GUSB mutations on GUSB expression and enzyme activity. HEK293T cells were transfected or not (NT) with an empty vector (EV) or a vector containing the cDNA encoding either the wild-type (WT) or the indicated mutant GUSB. Forty-eight hours after transfection, cells were harvested and cell lysates were prepared. (A) Protein expression of GUSB. Lysates (30 µg protein) were analyzed for GUSB expression by Western-blot. The blot is representative of three independent experiments. (B) Enzymatic activity of GUSB in transfected HEK293T cells. GUSB and Escherichia coli β-galactosidase activities were measured in cell lysates. The ratio of GUSB/E. coli β-galactosidase activities (means +/− SEM of three independent experiments) is presented as a percentage of the values recorded in cells overexpressing the WT GUSB. The basal enzyme activity of GUSB in non-transfected cells averaged 30 nmol/h/mg protein. Asterisks indicate statistically significant difference: *, p < 0.05; **, p < 0.01; ****, p < 0.0001.