Fig. 5: Sema can counteract the PI3K/AKT/Creb5 signaling pathway and reduce NR4a1 expression and mitochondrial-nuclear transport.

A Based on joint DEGs and Venn diagram analysis, network maps show the key molecular pathways involved in the cardiac remodeling process and regulated by Sema (n = 3). B Heatmap showing key molecular signatures involved in the cardiac remodeling process and regulated by Sema (n = 3). C Western blot analysis and quantification of PI3K, AKT, p-AKT, Creb5, NR4a1 and p-NR4a1 in the cardiac tissue of Sema-treated mice 8 weeks after sham or TAC surgery and quantification levels were normalized to β-actin (n = 6 independent experiments with similar results); PI3K/β-actin: F (3, 20) = 24.21, P = 7.3e-007; pAKT/AKT: F (3, 20) = 34.29, P = 4.5e-008; Creb5/β-actin: F (3, 20) = 17.38, P = 8.6e-006; NR4a1/β-actin: F (3, 20) = 6.550, P = 0.0029; pNR4a1/β-actin: F (3, 20) = 25.74, P = 4.5e-007. D, E Western blot analysis and quantification of NR4a1 in mitochondrial and nuclear proteins and quantification levels were normalized NR4a1 (nuclear/mitochondrial) (n = 6 independent experiments with similar results); NR4a1 (nuclear/mitochondrial): F (3, 20) = 5.807, P = 5.0e-003. F Pull-down analysis showing the binding between Creb5 and NR4a1 in vitro (n = 6 independent experiments with similar results). G, H Representative images of MIRO1 (green) and NR4a1 (red) immunofluorescence staining in NRVMs are shown on the left, and colocalization Pearson’s correlation coefficient (PCC) analysis is shown on the right (scale bar: 50 μm; n = 3 independent experiments with similar results). All results are shown as the mean ± SEM, and analysis using one-way ANOVA followed by Bonferroni post hoc test (C and E) was conducted. For the analysis in (G and H), a co-localization PCC analysis was used. p values are indicated. Source data are provided as a Source Data file. DEGs, differentially expressed genes; TAC, transverse aortic constriction; NRVMs, neonatal rat ventricular myocytes.