Fig. 2: Identification of CDK2AP1-antagonist miRs in OSCC cell lines.

A RT-qPCR expression analysis of the top 12 miR candidates arising from the microarray and in silico selection in CDK2AP1-deficient and -proficient cell lines. Of note, miR-592-5p expression levels are not shown due to undetectable expression. The dotted line indicates the average expression level of the analyzed miRNA in the proficient cell lines (CA1, LM, and LUC4). Experiments were performed in triplicate, and miRNA expression was normalized to that of snoRNA-U6. P values denote one-way ANOVA and one-sample t test against the mean of CDK2AP1-proficient cell lines (*P < 0.05; **P < 0.01; ***P < 0.001). B miR-21-5p RT-qPCR analysis of CDK2AP1-proficient cell lines upon transfection with either an empty (EV) or the pre-mir-21 PCDH-CMV lentiviral vector. Experiments were performed in triplicate and normalized to snoRNA-U6 expression. Fold change in miRNA expression was calculated relative to the EV-transfected cells for each experiment. One-sample t test P values: *P < 0.05; **P < 0.01; ***P < 0.001. C Western blot analysis of the consequences on CDK2AP1 and PDCD4 expression in the proficient cell lines transduced with an empty vector (EV) or with the pre-mir-21 PCDH-CMV lentiviral vector. Experiments were implemented in triplicate and β-actin (BACT) was employed as loading control. D Western blot analysis of the loss-of-function consequences of miR-21-5p inhibition on the CDK2AP1-deficient cell line panel. The SCCs cell lines were transfected with the MIRZIP lentiviral vectors targeting either miR-21-5p (mirZip-21) or a scrambled control sequence (miR-Zip-SCR). Experiments were performed in triplicate and β-actin (BACT) was employed as loading control.