Supplemental Material s18

SUPPLEMENTAL MATERIAL

Supplementary Figure 1. Analysis of miR-15a/16 effects in the PWC. NZB mice

(9mo) were injected with empty (GFP-Lentivirus, left) or mir-15a/16-lentivirus

(miR Lentivirus, right) and analyzed (8 days post) for GFP expression in B-1 cells

(CD5 dull, B220+), B-2 cells (CD5-, B220+) or T cells (CD5+, B220-). The percent GFP+ cells as a percent of the total in that population is indicated in the upper right corner of the histograms. GFP+ cells were found in the B cell population indicating that B cells preferentially were transduced. Supplementary Figure 2. Percent change in B-1 cells following short-term lentivirus treatment. NZB mice 9-12 mo were either uninjected or injected with mir-15a/16 lentivirus or control lentivirus and spleens analyzed 8 days post treatment. Columns are Mean % change ± SD. The % Change was calculated as

% B-1 in lenti - % B-1 untreated %B-1 untreated X 100

* indicates statistical significance, student’s t-test, p<0.05, n=6 control lenti, n=7 mir-lenti, n=3 untreated. Supplementary Figure 3. Effect of in vivo miR-15a/16 restoration in NZB mice on aneuploidy. NZB mice were injected with either a control lentiviral vector expressing GFP (control) only or a lentiviral vector expressing both GFP and the wild-type mir-15a/16-1 sequence (miR-15a/16). NZB were evaluated for short- term effects (8-9 days) and long-term effects (28-29 days) of miR-15a/16 restoration post-injection. A, Representative analysis of DNA content (by computer modeling) of NZB injected with control vector and miR-15a/16 lentiviral vector, (short-term = spleen, long-term = PWC). The first tallest peaks represent diploid G1 with the smaller peak to the right representing aneuploid G1. B,

Percentage of cells with aneuploidy in PWC, spleen, and PB from NZB injected with control or miR-15a/16-expressing lentivirus, left = short-term, right = long- term of PWC, spleen, and PB of NZB treated with control or miR-15a/16- expressing lentivirus. Black columns = control-lenti and white = miR-15a/16-lenti. Error bars represent SEM (short-term: control n = 5, miR-15a/16 n = 8; long-term: control n = 4, miR-15a/16 n = 4).

The percent of aneuploidy did not differ between NZB mice injected with either the control or miR-15a/16 lentiviruses in any tissue in the short-term study.

In the long-term study, which involved two injections, there was a trend in all tissues analyzed for a decrease in aneuploid cells in the long-term NZB injected with mir-15a/16-lentivirus when compared to NZB treated with control-lentivirus.