Fig. 4

Hes and Nar target the lipogenesis pathway. A Volcano plot representation of differentially expressed genes after Hes (left panel) and Nar (right panel) treatment. The x-axis indicates the expression fold change (FC) and the y-axis indicates the false discovery rate (FDR) (− log₁₀) for each gene versus untreated MM cells. FC threshold |1,5|; FDR ≤ 0.15. Upregulated transcripts (red) and downregulated (green) are shown. Selected representative genes are indicated with black arrows. B q-RT-PCR analysis of SREBF1 and c-MYC mRNA expression levels in AMO cells, 48 h after 250 µM Hes or Nar exposure. Histogram bars represent the average ± SD of mRNA expression levels after normalization with GAPDH and calculation using ΔΔCt method. *p < 0.05. WB analysis of C SREBP1, c-MYC D FAS, ACC, ACL and E DGAT2 protein levels, in AMO cells, 48 h after treatment with Hes or Nar. GAPDH or α-tubulin were used as loading control. F Fluorescence microscopy analysis of lipid droplets labeled with LipidTOX Red probe, 24 h after exposure to 250 µM of Hes or Nar. Representative images (100 × magnification) are reported. Histogram bars report the number of lipid droplets ± SD in at least 100 cells from three different fields. *p < 0.05. G Triglycerides measurement using Triglycerides Glo assay in AMO cells after 250 µM Hes or 250 µM Nar treatment. Luminescence was evaluated at 48 h. *p < 0.05