Fig. 6. miR-1271-5P interferes with FOXO1 expression during chondrogenic differentiation. (A) Heatmap of miRNA-seq analysis, showing the differently expressed miRNAs in the chondrogenic pellet differentiated from human iPSC-derived MSCs, and human iPSC-derived MSCs treated with DMSO or wedelolactone. (B) Volcano plots showing the differently expressed miRNAs. Red color indicates the significantly upregulated miRNAs, blue color indicates the significantly downregulated miRNAs, and gray color indicates the genes with no differential expression. (C) Venn diagram overlap showing the number of upregulated miRNAs and miRNAs in the DMSO group compared to the human iPSC-derived MSCs and wedelolactone groups. (D) Venn diagram overlap showing the numbers of downregulated miRNAs and miRNAs in the DMSO group compared to the human iPSC-derived MSCs and wedelolactone groups. (E) miR-1271-5P target sequence in the 3’-untranslated region (UTR) of FOXO1 predicted by the TargetScan database. (F) MiR-1271-5P target 3’-untranslated region (UTR) of FOXO1 was confirmed by the luciferase reporter assay. (G) Western blotting detection of protein expression levels of FOXO1 in human iPSC-derived MSCs transfected with the miR-1271-5P mimic or inhibitor under chondrogenic differentiation. (H) RT-qPCR analysis of miR-1271-5P in human iPSC-derived MSCs after EZH2 knockdown. Statistical differences were analyzed by one-way ANOVA followed by Dunnett’s test: *p<0.05, **p<0.01, ***p<0.001.

International Journal of Stem Cells 2023;16:326-41 https://doi.org/10.15283/ijsc22046

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