Figure 5
Obvious phenotypic changes in the morphology of Nrf1α−/− hepatoma cells.
(a) HepG2 (Nrf1α+/+) and HEA157 (Nrf1α−/−) cells were transfected with 1.5 μg of an expression construct for ER-DsRed marker protein. After the cells were allowed to recover for 12 h, subcellular location of Nrf1 was examined by immuocytochemistry with anti-Nrf1 antibody (the upper two rows, it should be noted that the anti-Nrf1 antibody is replaced by normal serum as an internal control in the third row), followed by confocal imaging. FITC-labelled second antibody was used to locate Nrf1 proteins. Nuclear DNA was stained by DAPI. The ER/DsRed gave a red image positively in the ER. The merge signal represents the results obtained when the three images were superimposed. (b,c) The above cells were subjected to observation of the morphological changes by light microscopy (b) and scanning electron microscopy (c), before relevant cell images were acquired. Overall, these images shown with different magnifications in sizes are a representative of at least three independent experiments undertaken on separate occasions that were each performed in triplicate (n = 9). The red arrowed cell was magnified to1500× than their original sizes (cf. right image with left image).
