Fig. 5: mTORC2-driven effects in epicardial maturation. | Nature Communications

Fig. 5: mTORC2-driven effects in epicardial maturation.

From: Generation of mature epicardium derived from human-induced pluripotent stem cells via inhibition of mTOR signaling

Fig. 5

a Immunocytochemical analysis of MKI67 expression in untreated, Torin1- and Rapamycin-treated hiPSC-epicardial cells at d24 + 7 (white arrows indicate positive signals; scale bar: 200 µm). Experiments were repeated with similar results 3 times. b Venn diagrams illustrating mTORC2-dependent elements in adult epicardium. Venn1 (top left) shows Torin1-specific transcription factors (TFs) not shared with Rapamycin. Venn2 (bottom left) presents TFs from differentially expressed genes (DEGs) significant in adult epicardium. Venn3 (right) displays the unique elements between Torin1-TFs (from Venn1) and adult-TFs (from Venn2). c mRNA of the top 5 upregulated TFs with Torin1 treatment. d Workflow for siRNA transfection against YBX3 and MAFF. Created in BioRender. Lucena-Cacace, A. (2025) https://BioRender.com/r4acodl. e mRNA expression of specific genes in Torin1-treated epicardial monolayers with siRNAs against YBX3 and MAFF (n = 4 biological replicates). Data are presented as means ± s.e.m.; two-tailed unpaired Student’s t-test. f Immunocytochemistry of siYBX3 Torin1-treated cells for WT1 and ZO-1 expression (scale bar: 100 µm). Experiments were repeated with similar results 3 times. g, h Western blot analyses of Torin1-treated hiPSC-epicardium with siRNAs against YBX3 and MAFF, examining proteins of the fetal program and cellular proliferation (WT1, TBX18, MKI67; β-actin normalization). Western blots were repeated with similar results 2 times. Source data is provided as a Source Data file.

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