Fig. 4: MiRNA signature impact on Androgen Receptor signaling by targeting WT1 and FOXA1. | Cell Death Discovery

Fig. 4: MiRNA signature impact on Androgen Receptor signaling by targeting WT1 and FOXA1.

From: Androgen receptor inhibition sensitizes glioblastoma stem cells to temozolomide by the miR-1/miR-26a-1/miR-487b signature mediated WT1 and FOXA1 silencing

Fig. 4

A Left panel: scatter plot of the Pearson’s correlation between the miRNA signature expression and AR in a TCGA miRNAseq dataset of 94 IDH wild-type glioma patients. Correlation coefficient (R) and linear trend line are reported; Right panel: Western blots of a representative experiment of AR expression in the indicated GBM neurospheres overexpressing the miRNA signature mimics or negative control mimic (neg. ctrl) after 6 days of TMZ treatment with the indicated doses. Densitometric analysis by ImageJ software is shown; B Schematic representation of AR inhibitors already evaluated on GBM; C IncuCyte time-course of tumor-spheres formation of TMZ- or ENZ-treated GBM cells after ectopic expression of the miRNA signature with respect to negative controls (neg. ctrl). Quantification of tumor-spheres diameter calculated by IncuCyte software; D Extreme limiting dilution analyses of TMZ- or ENZ-treated GBM cells after ectopic expression of the miRNA signature versus controls and graphic quantification of stem frequency. E mRNA expression levels of NANOG-, OCT4-, and SOX2- in TMZ-, or ENZ-treated GBM neurospheres overexpressing the miRNA signature compared to controls; F Tumor-spheres formation evaluated by IncuCyte time-course of TMZ- treated GBM cells after transfection of siRNAs- WT1 -FOXA1 or -AR versus controls. Quantification of tumor-spheres diameter calculated by IncuCyte software; G Left panel: Extreme limiting dilution analyses of TMZ- treated GBM cells after transfection of siRNAs- WT1 -FOXA1 or –AR versus controls and graphic quantification of stem frequency. Right panel: NANOG-, OCT4- and SOX2- mRNA expression levels in TMZ-treated GBM neurospheres after WT1, FOXA1, and AR downregulation by siRNA compared to controls. H Western blots of a representative experiment of MLKL and p-MLKL expression in TMZ-treated GBM neurospheres after WT1 and FOXA1 downregulation compared to controls. Densitometric analyses by ImageJ software are shown. *p ≤ 0.05, **p ≤ 0.01.

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