. 2010 Feb 13;19(10):1883–1896. doi: 10.1093/hmg/ddq064

Table 2.

Summary of human spastin transgenic lines and relative expression levels

Figure	Panel	Genotypes (line #) assayed	Relative spastin expression	Comments
2	B–D	H^WT(4),Ø	1X	Expression levels between these control genotypes (H^WT,Ø versus H^WT,H^WT) differ, but do not cause phenotypic consequences.
		H^WT(4),H^WT(6)	1X, 1X (=2X)
3	A and D	H^WT(4),H^WT(6)	1X, 1X	Expression levels are equivalent between genotypes. Differences are therefore due to a deleterious effect of R388.
		H^WT(4),H^R388(6)	1X, 1X
	B	H^R388(6),Ø	1X	Dosage may account for difference in degree of rescue conferred between R388 and STOP431.
		H^STOP431(9),Ø	0.5X
	C	H^WT(4),H^WT(6)	1X, 1X	One copy of H^WT rescues as well as two (see Fig. 2B). H^WT,H^R388 defects are therefore due to the R388 mutation rather than reduced expression.
		H^WT(4),H^R388(4)	1X, 0.5X
4	A	H^WT(4),H^WT(6)	1X, 1X*	*No differences observed despite 1X more expression in H^L44.
		H^WT(6),H^L44(6)	1X, 2X*
		H^L44(6),Ø	2X
		H^WT(4),H^R388(6)	1X, 1X**	**See Figure 3A and D comment.
		H^L44(6),H^R388(6)	2X, 1X***	***Significantly worse than H^WT,H^R388 despite 1X more of H^L44, which functions equivalently to H^WT and should therefore be alleviatory. Deleterious effects of R388 are therefore exacerbated by L44.
	B and C	H^WT(4 or 6),Ø	1X*	*No differences observed despite fold-differences in expression.
		H^WT(4),H^WT(6)	1X, 1X*
		H^WT(6),H^L44(6)	1X, 2X*
		H^WT(4),H^R388(6)	1X, 1X**	**See Figure 3A and D comment.
		H^L44(6),H^R388(6)	2X, 1X***	*** Significantly worse than H^WT,H^R388 despite 1X more of H^L44, which functions equivalently to H^WT and should therefore be alleviatory. Deleterious effects of R388 are therefore exacerbated by L44.
	D	H^WT(4),H^WT(6)	1X, 1X*	*Difference may be due to additional H^L44 expression or to the mutation itself.
		H^WT(6),H^L44(6)	1X, 2X*
		H^WT(4),H^R388(4) or H^WT(4),H^R388(6)	1X,0.5X or 1X,1X**	**Both R388 insertions yield significant differences versus H^WT,H^WT, regardless of reduced or equivalent expression levels, supporting a deleterious effect caused by the mutation.
		H^L44(6),H^R388(1) or H^L44(6),H^R388(6)	2X, N.D. or 2X,1X***	***Significantly worse than H^WT,H^R388 for all pair-wise combinations of recombinant lines, regardless of dosage (P < 0.001). Effects of R388 are therefore exacerbated by L44.
	E	H^WT(4),H^WT(6)	1X, 1X*	*L44 dose or mutation could underlie phenotype.
		H^WT(6),H^L44(6)	1X, 2X*	*L44 dose or mutation could underlie phenotype.
		H^WT(4),H^R388(6)	1X, 1X**	**Significantly different from H^WT,H^WT despite equivalent expression levels. Differences are therefore due to a deleterious effect of R388.
		H^L44(6),H^R388(6)	2X, 1X***	***Different dosage as H^WT,H^R388, but equivalent phenotype. Same dosage but different phenotype versus H^WT,H^L44. Phenotypes correlate with genotype and not dosage.
5	A and B	H^WT(4),Ø	1X	Reduced dosage of H^Q45 alone may account for differences from H^WT. However, H^Q45 still rescues as well or better than H^WT,H^R388, and exacerbates the effects of H^R388, consistent with genotype, and not dosage, underlying observed phenotypes.
		H^Q45(2),Ø	0.75X
		H^WT(4),H^R388(6)	1X, 1X
		H^Q45(2),H^R388(6)	0.75X, 1X

A total of 11 different transgenic lines expressing human spastin were used in these studies. Specific insertion lines (number denoted in parentheses) of the recombinant strains used in each experiment are listed, together with the estimated spastin expression level for each transgene, normalized to line 4 of H^WT (Supplementary Material, Fig. S1). Comparison between genotypes indicates that mutant phenotypes cannot be accounted for by differences in spastin dosage.

N.D., Not determined.