Figure 4. Arylsulfatase activity of media.
A: Arylsulfatase A (ARSA) products were analyzed with western
blot. ARPE-19 cells were transfected with 5 μg ARSA-FLAG and 1
μg SUMF1-FLAG, and cultured in serum-containing media for 2 h
and then in serum-free media for 22 h. Loaded was 12.5% of the total
sample. B: The ARS activity of ARSA-FLAG was measured using
0.4% of the total medium. C: Arylsulfatase I (ARSI) products
were analyzed with western blot. Transfected were 8 μg ARSI-FLAG, 8 μg ARSI-C93S-FLAG,
and 1 μg SUMF1-FLAG into ARPE-19 cells. D: The
trichloroacetic acid (TCA)-induced precipitate of the medium was
analyzed with western blot. E: The ARS activity of ARSI-FLAG
was measured using 20% of the total medium. F: The pH
dependence of ARS activities was shown. The ARS activities in B, E,
and F give the means of three independent experiments. Bars
indicate standard deviations.