Figure_1.tif (905.32 kB)

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

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posted on 2014-05-05, 04:06 authored by Maria De Salvo, Lavinia Raimondi, Serena Vella, Laura Adesso, Roberta Ciarapica, Federica Verginelli, Antonio Pannuti, Arianna Citti, Renata Boldrini, Giuseppe M. Milano, Antonella Cacchione, Andrea Ferrari, Paola Collini, Angelo Rosolen, Gianni Bisogno, Rita Alaggio, Alessandro Inserra, Mattia Locatelli, Stefano Stifani, Isabella Screpanti, Lucio Miele, Franco Locatelli, Rossella Rota

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.