Supplementary material for Redfern et al. (2000) Proc. Natl. Acad. Sci. USA 97 (9), 4826-4831.

The supplementary material for this article has been divided into four groups.

Group 1: Supplemental Methods and Fig. 7. The file (Methods and Fig. 7) is in PDF format and contains the following sections:

A: Methods Section

B: Ventricular Conduction Delay in a Mouse Expressing Ro1

C: Pertusis Toxin Reverts Wide QRS Complexes

D: Quantification of Collagen Formation in Ro1-Expressing Mice

E and F: Expression of Ro1 in Mouse Heart Decreases the Force and Speed of Myocardial Contraction

G: Echocardiograms of Control and Ro1-Expressing Mice

H: Drug-Induced Ro1 Signaling with Reduced Expression Levels

Group 2: Movie. This file is a QuickTime movie of the Ro1-induced cardiomyopathy, with recovery. Movie

Group 3: Data Sets. The following links provide gene expression data for more than 6,000 genes sorted by up- and down-regulation of Ro1 expression for 8 weeks. The compressed files expand into Microsoft Excel documents.

Data Set 1: Ro1-up-regulated genes

Data Set 2: Ro1-down-regulated genes

Ro1 DNA Array Analysis

The two data sets are in Microsoft Excel workbook format. Provided in these files are the averages, standard deviations, P values between control and experimental groups, and corresponding fold values for the 6,519 genes and expressed sequence tags (ESTs) present on the Affymetrix Murine6500 arrays.

The "8 weeks group" consisted of aMH-tTA/teto-Ro1 double transgenic mice (n = 9), raised on doxycyline (200 mg/kg in chow) from conception until adulthood. These animals were then taken off doxycycline and allowed to express Ro1 for 8 weeks until the time of death. The "control group" consists of aMH-tTA single transgenic mice (n = 8) littermate controls. All control mice were age- and strain-matched with the "8 weeks group" and raised on doxycycline from conception until death. (See the gene expression analysis section within the supplemental Methods section for all protocols.)

BLAST Definitions: Lists the Genbank BLAST definitions for all complete DNA sequences (cds) and ESTs. EST descriptions represent the highest ranking homologous sequence for the corresponding probe pair sets on the Murine6500 arrays.

Con Avg: Average gene expression values for the eight mice contained within the control group.

Con SD: Standard deviations for the average gene expression values for the eight mice contained within the control group.

8 wks Avg: Average gene expression values for the nine mice contained within the 8 weeks group.

8 wks SD: Standard deviations for the average gene expression values for the nine mice contained within the 8 weeks group.

p Ro1-8wks/Con: P value for the "8 wks Avg" versus the "Con Avg."

FoldRo1-8wks/Con: Fold values (from 1 to infinity and -1 to negative infinity) for the "8 wks Avg" versus the "Con Avg." Note: when calculating fold differences, all values less than 20 (including negative values) were rounded up to a value of 20. This step avoids unrealistic fold differences.

DatabaseAxnID: Genbank accession number.

Mu6500Gene Name: Affymetrix call name for oligoprobe sets on the Murine6500 arrays.

Group 4: Fig. 8. The following links contain two sets of biochemical pathway image maps for Ro1 gene expression data at 8 weeks. All of the genes contained upon these maps hyperlink directly to GenBank ENTREZ and Swiss-Prot references. Please see www.genmapp.org for more information.

Fig. 8A: G Protein Signaling

Fig. 8B: Inflammatory Response

Interactive Ro1 Pathways

Panels A and B of Fig. 8 are linked to an interactive web interface for G protein signaling pathways that are altered in the Ro1 expressing mouse model. The images contain HTML tags embedded in the graphics such that, when you click on a gene name (i.e., "AC5"), the description of that gene will appear.

The colored boxes in Fig. 8A represents gene expression data for mice expressing Ro1 for 8 weeks as compared to control mice. The boxes are colored according to fold and P value.

Behind each colored box in the Fig. 8A is a description of that gene or EST from the DNA array, GenBank, and SWISS-PROT database. To see these descriptions, click on the respective links for the "GenBank Accession #" or "SWISS-PROT ID." To see a description of the headers used for the DNA array gene expression data, as well as statistical methods, click on the link titled "see above description" under the header "Expression Profile" under each gene description entry.