Figure 1

Cyclin D1b and cyclin K are oncogenic in nude mice. a) Generation of LP-1-derived clones. Total proteins were extracted from individual clones, resolved by SDS-PAGE (12%) and immunoblotted with anti-cyclin D1 Ab which detects both cyclin D1a and b isoforms (DCS-6, BD Biosciences, Le Pont de Claix, France), anti-c-Myc Ab (sc-764, Santa Cruz Biotech., Santa Cruz, CA, USA), anti-Flag M2 Ab (Sigma-Aldrich, Saint Quentin Fallavier, France) which detects cyclin K construct. Anti-β-tubulin Ab (sc-9104, Santa Cruz biotech.) was used to control gel loading and transfer, GRANTA-519 cell line was used as control for cyclin D1a expression. The four clones then referred as LP-1cl1, LP-1D1b, LP-1 Myc and LP-1K marked with an asterisk (*) were injected in vivo. b) Each cell clone was injected with Matrigel s.c. in 5 nude mice which were sacrificed 8 weeks later. The number of mice with a tumor at the site of injection is presented in the histogram; two representative mice bearing tumor are shown as well as hematoxylin-eosin-safran (HES) staining of tumor sections.