Legends to the Supplementary Figures

Supplementary material for:

Wild-type apo A-I and apo A-IMilano gene transfer reduce native and transplant arteriosclerosis to a similar extent Yingmei Feng, Eline Van Craeyveld, Frank Jacobs, Joke Lievens, Jan Snoeys, Bart De Geest Journal of Molecular Medicine

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Supplementary Fig. 1. Representative Western blots showing the human apo A-I monomer band (28 kDa) at day 28 after gene transfer with 5 x 1010 particles of AdA-I and the human apo

A-I monomer and dimer bands (56 kDa) at day 28 after gene transfer with 5 x 10 10 particles of

10 AdA-IMilano or after transfer with 2.5 x 10 particles of AdA-I together with an equivalent dose of

AdA-IMilano. Supplementary Fig. 2. Distribution of HDL particles by size in AdA-I (Top panel) and AdA-

IMilano (Bottom panel) treated mice. Plasma was subjected to non-denaturing electrophoresis followed by human apo A-I Western blot. Computer assisted scanning densitometry using co- electrophoresed molecular size calibrators was used to determine lipoprotein particle sizes. Supplementary Fig. 3. Cholesterol lipoprotein profiles obtained by fast performance liquid chromatography gel filtration of 100 µl plasma at day 0 (), day 14 (), day 35 () and day 70

10 () after transfer with 5 x 10 particles of Adnull (A), AdA-I (B) or AdA-IMilano (C) or after

10 transfer with 2.5 x 10 particles of AdA-I together with an equivalent dose of AdA-IMilano (D) in male C57BL/6 apo E-/- mice. Fractions 12-32 correspond to non-HDL sized particles whereas fractions 33-45 correspond to HDL sized particles.

Legends to the supplementary tables

Supplementary Table 1. Human apo A-I monomer and dimer concentration at day 28 after gene

10 10 transfer with 5 x 10 particles of AdA-I or AdA-IMilano or after transfer with 2.5 x 10 particles of

-/- AdA-I together with an equivalent dose of AdA-IMilano in male C57BL/6 apo E mice.

AdA-I AdA-IMilano AdA-I/AdA-IMilano

Monomer 230 ± 38 76 ± 11* 95 ± 9.2*

Dimer 0 ± 0 190 ± 22 150 ± 26

Data are expressed in mg/dl and represent means ± SEM (n=6). *: p<0.05 versus AdA-I. Supplementary Table 2. Immunohistochemical analysis of allograft inflammation at day 21 after transplantation. Data are expressed in percentages and represent means ± SEM.

Controls AdA-I AdA-IMilano Number of mice 14 16 12 Leukocytes 60 ± 2.3 44 ± 4.4** 49 ± 2.6* Macrophages 49 ± 2.6 31 ± 3.7*** 38 ± 2.1* T-lymphocytes 8.0 ± 1.1 5.6 ± 1.0 8.0 ± 1.4 *: p<0.05; **: p<0.01; ***: p<0.001 versus Controls.