Fig. 3.

Sdc4-Tg mice show increased NFAT activation upon pressure overload. A Relative NFAT-luciferase (luc) activity in whole left ventricle (LV) of Sdc4-Tg -NFAT-luc and NFAT-luc mice at baseline (6–8 weeks of age, n = 8), and after two weeks of aortic banding (AB) or sham operation (n = 6–15). See Table SII for mouse characteristics. Relative mRNA levels of calcineurin (encoded by Ppp3cb; B) and nuclear factor of activated T-cells c4 (NFATc4, encoded by Nfatc4; C) in the LV of Sdc4-Tg and WT mice after 20 weeks of AB. Gene expression was normalized to Gapdh. Data are mean ± SEM. Statistical differences were tested using t-test (baseline), or one-way ANOVA with Dunnett´s post-testing vs. NFAT-luc sham, *p < 0.05; and vs. NFAT-luc AB2w, ^#p < 0.05 (A), or using one-way ANOVA with Dunnett´s post-testing vs. respective WT sham, ***p < 0.001; and vs. WT AB20w. D Immunoprecipitation of full-length (FL) syndecan-4 from LVs of Sdc4-Tg mice, using an antibody detecting an extracellular epitope (see Fig. S1 for syndecan-4 antibody validation), or rat IgG as control. Representative immunoblots of syndecan-4 and calcineurin (n = 3). As positive controls for the calcineurin antibody, mouse brain lysate and recombinant calcineurin (130 kDa) was used. As positive control for the syndecan-4 antibody, overexpression of hemagglutinin (HA)-tagged syndecan-4 in HEK293 cells was used as positive control