Fig. 4

Bioinformatics data analysis shows that the gene most associated with the heterogeneity of MSCs in regulating Tregs differentiation is TNFSF4. (a) Bubble plot showing the GO enrichment analysis of immune-related significant pathways of DEGs between UCMSCs and DMSCs, especially enriched in plenty of T cell related pathways. (b) Gene expression heatmap showing the expression levels of 21 Tregs-related genes between UCMSCs and DMSCs. The color of each square indicates the scaled expression level of each gene. The columns show each sample, while the rows show each DEGs related to Tregs. (c) Bar plot indicates the Tregs-related DEGs between UCMSCs and DMSCs. There was significant difference for TNFSF4 with the maximal foldchange. Compared to DMSCs, the orange represents Tregs-related genes that are upregulated in UCMSCs, while the blue represents those that are downregulated. (d) The expression levels of TNFSF4 in UC-Mix and D-Mix were determined by qPCR. (e-f) The proportion of TNFSF4⁺ cells in the UC-Mix and D-Mix were determined by flow cytometric analysis. (g-h) Protein levels of TNFSF4 in UC-Mix and D-Mix were determined by western blotting and quantified. Images of unedited full blots in Figure S9