Fig 1: Enzyme's physical levels in the liver over time.Mice were hydrodynamically injected into the tail vein with 50 µg of plasmids encoding either IDUA or LT-IDUA. Enzyme physical levels in the liver were measured with ELISA at ten timepoints ranging from 0.5 to 168 h post-injection. Each data point represents one mouse. Data points from 8 to 168 h post-injection are shown in this figure, as time points earlier than 8 h yielded absorbance readings outside the standard curve.
Fig 2: Enzyme activity levels over time in liver.Mice were hydrodynamically injected into the tail vein with 50 µg of plasmids encoding either IDUA or LT-IDUA, and enzyme activity levels were measured in the liver at ten timepoints ranging from 0.5 to 168 h post-injection. Each data point represents one mouse, and the line is the mean enzyme activity for a timepoint. Standard deviation bars are not shown in the figure because each timepoint had three to four mice per group. The lower limit of quantification for the noncompartmental analysis was set as 2.198 nmol/h/mg.
Fig 3: Enzyme activity levels in the tissues and plasma.Mice were hydrodynamically injected into the tail vein with 50 μg of plasmids encoding different iduronidase enzymes (IDUA, p12-LS-IDUA, p12-LT-IDUA, LS-IDUA, and LT-IDUA). The IDUA group served as a comparator group for the four modified IDUAs. After 48 h post-injection, iduronidase levels were measured in the tissues and plasma using an enzyme activity assay. Each data point represents one mouse, and the mean ± 95% confidence interval for each group is shown. A one-way ANOVA with multiple comparisons adjustment was performed, and statistically significant differences between plasmid-treated groups are shown.
Fig 4: No Relationship Between Enzyme's Physical Levels in the Liver and Enzyme's Activity Levels in the Liver.Mice were hydrodynamically injected into the tail vein with 50 µg of plasmids encoding either IDUA or LT-IDUA. Enzyme levels in the liver were measured with enzyme-linked immunosorbent assay (ELISA) and an enzyme activity assay at timepoints ranging from 8 to 168 h post-injection. Each data point represents one mouse. In the IDUA group, there was no correlation between the enzyme's physical levels in the liver and the enzyme's activity levels in the liver (IDUA r2 = 0.05341, p = 0.5206). Similarly, in the LT-IDUA group, there was no correlation between the enzyme's physical levels in the liver and the enzyme's activity levels in the liver (LT-IDUA r2 = 0.01666, p = 0.7223).
Fig 5: No relationship between age at injection and enzyme activity levels.Mice were hydrodynamically injected into the tail vein with 50 μg of plasmids encoding different iduronidase enzymes (IDUA, p12-LS-IDUA, p12-LT-IDUA, LS-IDUA, and LT-IDUA). Iduronidase levels were measured in the heart and plasma using an enzyme activity assay after 48 h. Pearson's correlation tests were performed for age at injection and enzyme activity levels. (A) Enzyme activity levels in the heart did not correlate with age at injection (r2 = 0.01640, p = 0.4190). (B) Enzyme activity levels in the plasma did not correlate with age at injection (r2 = −0.04050, p = 0.2013).
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