Fig. 2

Tregs promote the proliferation of neonatal CMs. a Rat neonatal CMs were exposed to the supernatant (SN) conditioned by Tregs and stained for various proliferation markers, including EdU incorporation, Ki67, histone H3 phosphorylated on serine10 (H3PS10), and Aurora B. All proliferation markers are stained red, while nuclei are stained blue with DAPI and CMs green using anti-α-actinin antibodies. Control cells were exposed to the medium normally used to culture Tregs, containing recombinant IL2 (50 ng/ml). Scale bar, 25 μm. b–e. Quantification of EdU⁺ (b), Ki67⁺ (c), H3PS10⁺ (d) cells, and AuroraB⁺ localization in midbodies (e) in either CMs (CMs, white bars) or fibroblasts (FBs, gray bars), expressed as the percentage of the total number of nuclei belonging to each cell type. All values are mean ± s.e.m., n = 3 biological replicates. At least 500 cells were analyzed per each replicate using the MetaXpress software. Pairwise comparison was performed with the Student’s t-test (b–e); *P < 0.05, **P < 0.01, ***P < 0.001 relative to control