Forced expression of Notch3IC in RH30 and RD RMS cell lines enhances cell proliferation and soft-agar colony formation <i>in vitro</i>.

<p>A, cell proliferation was assessed on RH30 and RD cells stably transfected with Notch3IC-expressing plasmid (RH30-Notch3IC and RD-Notch3IC) or with an empty control plasmid (RH30-Vector and RD-Vector). Representative of three independent experiments in duplicate (*P<0.05); Bars, SD. B, Western blotting showing the levels of the intracellular active form of Notch3 (Notch3IC), HES1, p21Cip1, phosphorylated ERK1/2 (p-ERK1/2), total ERK1/2 (ERK1/2), phosphorylated Akt (p-Akt), total Akt (Akt), phosphorylated p38MAPK (p-p38), p38MAPK (p38) and Myogenin in RH30 and RD cells stably transfected with either a pcDNA3 expressing Notch3IC (Notch3IC) or an empty pcDNA3 vector as control (Vector). Actin was the loading control. C, 30000 living cells (evidenced by trypan blue exclusion) stably over-expressing Notch3IC (RH30-Notch3IC and RD-Notch3IC) and their control (RH30-Vector and RD-Vector) were seeded on soft-agar in 35-mm Petri dishes and the number of colonies per field were quantified under a light microscopy.