Fig. 1: NRF2 expression and activity are strongly downregulated in different glioblastoma cellular models silenced for Caspase-8 expression.

A Bar chart of top enriched downregulated terms derived from the ENCODE_and_ChEA_Consensus_TFs_from_ChIP-X gene set library by using EnrichR Tool. The top 10 enriched terms for the input gene set (GSE193495) are displayed based on the -log10(p-value), with the actual p-value shown next to each term. The term at the top, NFE2L2, has the most significant overlap with the input query gene set. B Real time-PCR of NRF2 gene in U87-MG, U251 and GBMSC83 shCTR and stably silenced for Caspase-8 expression (U87-MG and U251 using shC8#1 sequence, GBMSC83 using shC8#2 sequence) (actin: housekeeping gene). C Immunoblotting of NRF2 and Caspase-8 in U87-MG, U251 and GBMSC83 shCTR and stably silenced for Caspase-8 expression (shC8#1 or shC8#2). GAPDH was used as loading control. D Densitometric analysis of NRF2 in (C) normalized on GAPDH. E Representative immunofluorescence of U87-MG, U251 and GBMSC83 shCTR and shC8 cells. NRF2 (green) and DNA (Hoechst, blue) and F relative histograms of the NRF2 staining reported as the ratio between nuclear and cytosolic fluorescence intensity (N/C). G Real time-PCR of NRF2 target genes (sqstm1, gclc, hmox1, nqo1) in U87-MG, U251 and GBMSC83 shCTR and shC8 cells (actin: housekeeping gene). Results represent the mean of at least three independent experiments ( ± SEM). Statistical analyses: paired (B, D, F), and multiple (G) t test (*P < 0.05; **P < 0.01; ***P < 0.001).