Fig. 3

miR-101-3p directly targeted LMNB1. A Western Blot analysis of LMNB1 protein levels in MEL-JUSO and SK-MEL-28 transfected with miR-101-3p mimic and siCtrl, respectively (Student’s t-test). B Immunofluorescence staining of protein expression of LMNB1 in MEL-JUSO and SK-MEL-28 treated with miR-101-3p mimic (72 h) and respective siCtrl. Representative images of LMNB1 expression (red) in MEL-JUSO and SK-MEL-28 transfected with miR-101-3p mimic and siCtrl, respectively. Nuclear staining with DAPI (blue). Scale bars, 20 µm (Student’s t-test). C Luciferase assay of a pGL3prom construct with a miR-101-3p binding site for LMNB1 in the 3′-UTR and the corresponding mutagenesis constructs, revealing LMNB1 as a direct target of miR-101-3p (one-way ANOVA and subsequent Fisher’s LSD multiple comparison test). Bars show mean ± SEM (*p ≤ 0.05, ns not significant)