Fig. 2

Stable aggregate formation is associated with miR-10b expression. a DIG-labelled miR-10b-5p specific LNA probes (green) were used in an in-situ hybridization protocol to quantify relative expression of miR-10b in cultured cells by FITC labeled anti-DIG-Fab immunofluorescence imaging. All three tested RDEB-cSCCs and two out of three HC-cSCC cell lines demonstrated increased miR-10b signals compared to primary HC-KCs. b To quantify the miR-10b expression at single cell resolution, cytoplasmic FITC signals of cells were cumulated and plotted as relative fluorescence intensity per single cell on the x-axis. Graphic shows distribution density function of miR-10b intensity of all analyzed single cells per sample. Respective negative- (scrambled) and positive- (U6) probes were used as controls (Supplementary Figs. S3 and S4 in Additional File 1). c All RDEB-cSCC lines and two out of three HC-cSCC lines demonstrated stable aggregate formation 96 h post seeding (representative images of aggregates). d Cross section of objects evaluated after transferring aggregates (n~ 7·10³) into cell culture plates. Each dot represents one object. e In addition, overexpression of miR-10b in KCs conferred spheroid formation capacities. Sample codes HC-KC: 1090KC, RDEB-KC: RDEB-57KC. Scale bars: 50 μm (white), 100 μm (black)