Figure 7
Increases in the migration and invasion of Nrf1α−/− cells and their clone formation on soft agar.
(a–d) HepG2 (Nrf1α+/+) and HEA157 (Nrf1α−/−) cells were starved for 12 h in a serum-free medium and then subjected to transwell migration (a) and invasion (c) assays as described in the section of ‘Materials and methods’. The migratory and/or invasive cells, that had passed through the 8-μm microporous membrane and attached to the lower surface of the transwell membranes, were fixed with 4% paraformaldehyde and stained with 1% crystal violet reagent before being counted. The results were calculated as a fold change (mean ± S.D.) of migratory (b) and invasive (d) Nrf1α−/−cells, which are shown as a representative of at least three independent experiments undertaken on separate occasions that were each performed in triplicate. Significant increases ($p < 0.05, n = 9) are indicated, relative to the corresponding control values obtained from wild-type Nrf1α+/+ HepG2 cells. (e,f) The soft agar colony formation of the above two cell lines was examined as described in the text of ‘Materials and Methods’. The resulting cell clones formed on the soft agar plates were stained with 1% crystal violet reagent before being counted. (f) The data were calculated as a fold change (mean ± S.D.) of the number of Nrf1α−/− cell clone formation and the significant increase ($p < 0.05, n = 9) is analyzed, relative to the control values of Nrf1α+/+ cells.
