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Fig. 3 | Cell Communication and Signaling

Fig. 3

From: A cancer stem cell-like phenotype is associated with miR-10b expression in aggressive squamous cell carcinomas

Fig. 3

Knock-out of MIR10B reduces aggregate sizes. a, b Knock-out of MIR10B in RDEB-cSCC shifts the distribution of cell aggregates towards an increased number of single cells and aggregate fragments (indicated as small objects) in a size distribution analysis by cross-section of formed aggregates compared to parental cells. This effect was reversed in a mixed culture of knock-out and parental cells. c The significant difference in size distribution was evaluated by a Kolmogorov-Smirnoff (KS) test, where the null distribution of the KS test statistic was derived by Monte Carlo simulation upon random resampling (parental, MIR10B−/− and mixed). d Extended culture period over several passages (> 10 passages) of MIR10B knock-out cells led to the prevailance of residual miR-10b expressing cells, resulting in a mixed population of RDEB-SCC1MIR10B−/− and parental miR-10b expressing cells. In addition, upon short trypsinization of a mixed population, we found that MIR10B knock-out cells detached earlier, whereas the remaining cells, showed an enrichment of those cells still harboring the genomic MIR10B locus. (PCR on MIR10B locus in isolated genomic DNA). e Proliferation of miR-10b expressing parental cells showed no difference to MIR10B knock-out cells or to a mixed population. Proliferation was assessed over 7 days. Mean ± SEM are given, n = 4 ind. repeats. f Spheroids formed by miR-10b expressing parental RDEB-cSCC cells and a mixed population showed outgrowth of cells 24 h post aggregate transfer to cell culture plates. This phenomenon was much less pronounced in MIR10B knock-out cells. HC-cSCC (e.g. SCC13) also presented with outgrowth from spheroids within 24 h post transfer similar to RDEB-cSCC (representative images from two independent repeats)

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