Figure 5: In vitro residual sulfatase activity and FGE stability of novel SUMF1 variants.
(A) Steroid sulfatase (STS) and arylsulfatase A (ARSA) activities were measured in cells harbouring each of the 9 novel SUMF1 variants. Bar diagram depicting the comparison of the specific activity of STS and ARSA relative to activity of cells expressing FGE-WT (Also see Supplementary Fig. S1). The values represent mean ± SEM of three independent experiments. (B) The stability of the novel FGE variants were compared to wildtype and the half-life (t1/2) was calculated (see legend). Plot depicts the percentage of FGE remaining after cycloheximide chase for the indicated time points. The amount of FGE was determined by quantification of signals corresponding of FGE in western blots (See Supplementary Fig. S2). After normalization of the anti-HA antibody signals (corresponding to FGE) in cell lysates to anti- Hsc70 signals, the total amount of FGE (μg/mg of total protein in lysate) in the cells and media were combined and expressed as the percentage of that at the start of the chase (0 h).