Fig 1: Metabolic variations in the course of NB4 differentiation along the myeloid lineage after treatment with ATRA. (A) WikiPathways analysis of the glycolytic pathway enriched in ATRA-regulated genes (FC|1.5|; FDR < 5%). The Transcriptome Analysis Console (TAC) 4.0.2 (Affymetrix, Inc.) allowed to observe that the “metabolic reprogramming pathway” was significantly enriched (p < 0.001; Fisher’s exact test) in ATRA-regulated genes (nine upregulated genes, and one downregulated gene). Red and green boxes represent up and downregulated genes, respectively. (B) WikiPathways analysis of OXPHOS system enriched in ATRA-regulated genes (FC|1.5|; FDR < 5%). The “metabolic reprogramming pathway” was found significantly enriched (p < 0.001; Fisher’s exact test) in ATRA regulated genes (10 upregulated gene, and 7 downregulated genes). Red and green boxes represent up and downregulated genes, respectively. (C) Validation of some differentially expressed glycolytic and OXPHOS enzymes in NB4 cells treated with 1 µM ATRA for 120 h in comparison with untreated NB4 cells. Representative immunoblot analysis of Aldolase C, LDHB, NDUFB6, PGAM1/4, and SDHA proteins. GAPDH and ß-Actin were used as loading controls. (D) Quantification of immunoblot experiments. Data are reported as mean ± SD of at least three independent experiments (Student’s t-test, ** p < 0.01; *** p < 0.001, with respect to relative controls; a.d.u., arbitrary densitometric unit).