Figure 5.
tNOX overexpression in NIH3T3 cells enhances cell growth. NIH3T3 cells were transfected with GST, GST–tNOX wild-type, GST–tNOXS504D, or GST–tNOXS504A. (A,B) Overexpression of GST–tNOX wild-type, GST–tNOXS504D, or GST–tNOX S504A fusion proteins was analyzed using anti-GST, anti-tNOX, anti-phospho-ERK, and anti-ERK antibodies. β-Actin was detected as a loading control. Representative images are shown. (C) The cell-doubling time was determined by CMFDA staining of NIH3T3 cells. The presented values (mean ± S.E.) represent at least three independent experiments (** p < 0.001). (C) Dynamic monitoring of cell proliferation was performed using impedance technology, as described in the Materials and Methods section. Normalized cell index values measured over 138 h are shown.