Figure 3

Hemin modulates tumour suppressor proteins. (A) Hierarchical cluster analysis shows the significant differences (*p < 0.05; Student's T-test) in the expression of tumour suppressor genes detected in LM3 cells in control- versus hemin-treatment. The Z-score of median MS intensities (n = 7) are colour coded to show relative protein abundance. Shown in red are the up-regulated proteins and shown in blue are the down-regulated proteins after hemin treatment. The bar below indicates how many times the expression increased or decreased (Z-score). Western blot corroborates the protein expression levels of Smad2/3, PTEN (B) and p53 (C). GAPDH was used as loading control. Bottom: densitometric quantification of each protein (n = 3), (*p < 0.05; Student's T-test). (D) Immunohistochemistry of Smad2/3 and PTEN in LM3 syngeneic mouse model biopsies showing the differences in staining between vehicle- and hemin-treated mice. (E) Immunoreactive score (IRS) quantification of Smad2/3 and PTEN protein levels (n = 4–5) (*p < 0.05; Student's T-test).