Figure 1

Effects of hemin treatment in LM3 breast cancer cells proteome. (A) LM3 cells were treated with 80 µM hemin or vehicle (control) for 24 h. Plot profile of label free quantification (LFQ) intensity of HO-1 protein in control (n = 7) versus hemin treatment (n = 7) is shown. (B) Western blot analysis shows the expression of HO-1 protein. GAPDH was used as loading control. Bottom: Bar graph shows the densitometric quantification of HO-1/GAPDH ratio (n = 3) (ImageJ quantification). (C) Venn diagram of the total number of proteins differentially expressed with hemin respect to the control. After hemin treatment 1309 proteins were up regulated (red), 921 were down regulated (blue) and 4147 were without any change (*p < 0.05; Student's T-test). (D) Heatmap shows the Z-score of significant LM3 protein levels (Student's T-test, S0: 0.5; Permutation-based FDR: 0.05) showing main protein clusters with or without hemin-treatment. The colour code shows in red the up regulated proteins, in blue the down regulated ones and in white the proteins with no significant variation between the vehicle- and hemin-treated cells. Gene ontology-terms enriched in the MS proteome and STRING network analysis showed the clusters of hemin up-modulated proteins (red circles) and the most down-regulated proteins after hemin treatment (blue circles). The functional protein-associated networks show the interconnection between modulated proteins and the edges indicate both functional and physical protein associations.