CD271 modulation affects cSCC spheroid phenotype a Representative picture of in situ, WD, or MD/PD patient-derived CD271-transduced or mock spheroids. Scale bar = 100 μm b CD271 expression by qPCR. β-actin was used as housekeeping gene. c Spheroid total area measured by ImageJ software. d Spheroid viability evaluated by MTT assay. e E-cadherin, KRT10, and Slug expression evaluated in patient-derived spheroids by western blotting. Tubulin was used as loading control. f Representative picture of SCC13 spheroids treated as described. Scale bar = 100 μm g Spheroid total area measured by ImageJ software. h Spheroid viability evaluated by MTT assay. i CD271, E-cadherin, KRT10, ERK1/2, pERK1/2, and Slug expression evaluated in CD271-overexpressing or silenced SCC13 spheroids by western blotting. pERK1/2/ERK1/2 expression ratio analyzed by ImageJ software. β-actin was used as loading control. j GO Biological Process term fold-Enrichment of RNA-seq data of CD271-transduced SCC13 spheroids vs mock determined by PANTHER Tools (a triplicate for n = 80 mock or CD271 spheroids). For all experiments, the results are shown as mean ± SD of three independent experiments. Statistical analysis was performed using the two-way ANOVA or multiparametric T-test. *0.01 < p < 0.05, **0.001 < p < 0.01, ***0.0001 < p < 0.001, ****p < 0.0001
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