Figure 1.
Immunolocalization of Matn2 in the liver of young mice (A).
Matn2 immunostaining is most intense around the portal blood vessels (arrowhead) on a frozen section of a 40-day old WT mouse liver. Matn2 partially colocalizes with laminin; however, in several structures Matn2 staining is more intense (arrowheads). There is no immunosignal in the matching area in the liver of a Matn2-/- mice. Bar, 0.1 mm. Representative histological stain of WT control, WT DEN-treated, Matn2-/- control and Matn2-/- DEN-treated mouse livers (B). Scale bars represent 0.1 mm.
Figure 2.
Representative immunohistochemical stains of WT control, WT DEN-treated, Matn2-/- control and Matn2-/- DEN-treated mouse livers (A).
Scale bars represent 0.1(insets) for paraffin-embedded samples. Changes in cell cycle regulation. Ki-67 proliferation index (B). In knockout samples an average of 4.1 Ki-67 positive cells were counted per field of view compared to 0.7 in WT (p<0.001) (B). Results are expressed as mean ± SD. Representative Western blots of cell cycle regulatory proteins (C) in WT control, WT DEN-treated, Matn2-/- control and Matn2-/- DEN-treated mouse livers. Diagrams of band intensities expressed as values normalized to β-Actin loading control (D). Data are expressed as mean ± SD, n = 3.
Figure 3.
Representative Western blots of intracellular regulatory proteins
in WT control, WT DEN-treated, Matn2-/- control and Matn2-/- DEN-treated mouse livers (A/1, A/2). Results of densitometrical analysis of band intensities expressed as values normalized to β-Actin loading control (B). Data are expressed as mean ± SD, n = 3.
Figure 4.
Representative immunofluorescence and immunohistochemical stains of WT control, WT DEN-treated, Matn2-/- control and Matn2-/- DEN-treated mouse livers.
Scale bars represent 0.1(insets) for paraffin-embedded samples and 0.05 mm for frozen tissue sections.
Figure 5.
Results of Phospho-RTK antibody array.
Picture of RTK array membrane (A). Densitometry of phosphorylation signals in WT control (dark gray bars) and WT DEN-treated (white bars) samples, compared to control (black bars) and Matn2-/- DEN-treated (light gray bars) samples (B). Data are expressed as mean ± SD, n = 3.
Table 1.
Differences between WT control and Matn2-/- control group compared to WT control.
Figure 6.
Results of DEN treatment in WT and Matn2-/- animals.
Representative pictures of the macroscopic appearance of WT and Matn2-/- DEN-treated mouse livers. 10 months after DEN exposure the number and size of macroscopic tumors were greater in Matn2-/- than in WT livers (A). The number of tumors/animal (B) and the tumor volume (C) were significantly higher in Matn2-/- mice compared to WT mice after DEN-treatment (n = 9 for WT DEN, n = 13 for Matn2-/- DEN, **p<0.01).
Table 2.
Differences between WT control and WT DEN group compared to WT control.
Table 3.
Differences between Matn2-/- control and Matn2-/- DEN group compared to Matn2-/- control.
Table 4.
Differences between WT DEN and Matn2-/- DEN group compared to WT DEN.
Figure 7.
Schematic illustration of the predicted signaling mechanisms in Matn2-/- mice.
Molecules studied in this work are labeled by asterisks. Empty boxes show upregulation and/or activation; gray boxes indicate downregulation and/or inactivation of the protein.