Fig. 4

Reexpression of miR-101-3p and its influence on apoptosis and cell cycle. A Flow cytometry analysis for apoptotic cell detection in miR-101-3p mimic-transfected MEL-JUSO, SK-MEL-28, and MV3 (72 h) compared with siCtrl using Annexin V-FITC and PI staining (Student’s t-test). The pictures indicate one exemplary staining of MEL-JUSO, SK-MEL-28, and MV3. B Flow cytometry with the fluorescent dye propidium iodide for cell cycle staining of sub-G1 phase. MEL-JUSO, SK-MEL-28, and MV3 transfected with miR-101-3p mimic (72 h) and siCtrl, respectively (Student’s t-test). C Flow cytometry with the fluorescent dye propidium iodide for cell cycle staining of G1, S, and G2 phase. MV3 transfected with miR-101-3p mimic (72 h) and siCtrl, respectively (two-way ANOVA and subsequent Fisher’s LSD multiple comparison test). D Cell cycle analysis with the FUCCI reporter system in MV3 cells transfected for 72 h with miR-101-3p mimic and the respective siCtrl. Representative fluorescence staining with bright-field overlay of MV3 FUCCI miR-101-3p mimic- and siCtrl-transfected cells. Scale bars, 20 μm (two-way ANOVA and subsequent Fisher’s LSD multiple comparison test). E–G GSEA enrichment plots of the gene sets Hallmark Apoptosis, GOBP of Mitotic G1-S Transition Checkpoint Signaling, and Hallmark P53 Pathway (FDR < 25%), illustrating the profile of the running enrichment score (green) and positions of the enriched gene set and the rank-ordered list of genes differentially expressed in MEL-JUSO treated with miR-101-3p mimic and siCtrl, respectively. Genes upregulated in miR-101-3p mimic melanoma cells are shown on the left side of the graph in red, and downregulated ones on the right side in blue. Bars show mean ± SEM (*p ≤ 0.05, ns not significant)