Fig 1: ATRA effects on the expression of the genes belonging to the glycerophospholipid metabolism pathway in parental NB4, NB4-shCTRL, and NB4-shATG5 cells.A The effects exerted by ATRA on the genes belonging to the glycerophospholipid pathway in parental NB4, NB4-shCTRL, and NB4-shATG5 cells were evaluated using the RNA-seq data generated from the three cell lines exposed to vehicle (DMSO) or ATRA (1 µM) for 48 h. The diagrams show the perturbations induced by ATRA on the expression of the indicated genes. The results presented in each box are expressed using the Log2 of the ATRA-dependent fold-change values which we determined. The names of the genes whose expression is significantly increased by ATRA in the indicated cell-lines are written in red. The names of the genes whose expression is significantly decreased by ATRA in the indicated cell-lines are written in blue. When the indicated genes are significantly up-regulated (FDR > 0.05), the positive fold-change values are contained in a red box and are marked with an asterisk. When the indicated genes are significantly down-regulated (FDR > 0.05), the negative fold-change values are contained in a light blue box. B The column diagrams indicate the expression levels of the CRLS1, LPCAT1, and PGS1 mRNAs which were determined in three independent cultures of NB4-shCTRL and NB4-shATG5 cells, by PCR using specific Taqman Assays. The results are expressed as the Mean + S.D. (N = 3) of the values determined. **Significantly different relative to the corresponding vehicle-treated control (p < 0.01, unpaired Student’s t test). The diagrams shown are representative of two independent experiments producing similar results. C Western blot analyses were performed on pooled cellular extracts obtained from three independent cultures of parental NB4, NB4.306, NB4-shCTRL, and NB4-shATG5 cells exposed to vehicle (DMSO) or ATRA (1 µM) for 48 h. The Western blots were performed with the use of specific antibodies targeting the indicated proteins. The same amounts of proteins were loaded in each lane, as demonstrated by the Western blot signals obtained with the anti-ß2actin (ACT) antibodies. The Western blots shown are representative of two independent experiments producing similar results. D The Western blots summarize the effects of ATRA on the expression of CRLS1 and LPCAT1 proteins in the cytosolic and mitochondrial fractions of NB4 cells exposed to vehicle or ATRA. Three independent cultures of NB4 cells were treated with vehicle (DMSO) or ATRA (1 µM) for 48 h. At the end of the treatment the mitochondrial and cytosolic fractions of the cell homogenates were isolated. The mitochondrial and cytosolic fractions were pooled and subjected to Western blot analysis with antibodies targeting CRLS1, LPCAT1, PGS1, cytochrome-c (CYT-C) and actin proteins, as indicated. The same amount of cytosolic and mitochondrial proteins was loaded in each couple of lanes (vehicle and ATRA). The Western blots shown are representative of two independent experiments producing similar results. E The scheme summarizes the effects exerted by ATRA in APL blasts, which lead to terminal granulocytic maturation of the NB4 cells.