Figure 2.

RanBP9 and TSSC3 function cooperatively to inhibit anchorage-independent cell growth and sensitize osteosarcoma cells to anoikis. SaOS2 or MG63 cells were stably transfected with the indicated constructs. (a and b) Soft agar assays. SaOS2 (a) or MG63 (b) cells (20 000) were seeded, cultured for 10 days in media supplemented with 10% FBS and the number of colonies was determined. (c, and d) Apoptosis assays. SaOS2 (c) or MG63 (d) cells were cultured under suspension conditions and subjected to flow cytometric analysis. Representative flow cytometric analyses (i) and quantification (ii and iii) of the percentages of early apoptotic cells (Annexin V⁺, PI^-) and late apoptotic or necrotic cells (Annexin V⁺/PI⁺). Note: all values are mean±S.D. of three independent experiments each performed in triplicate; *P<0.05, **P<0.01, ***P<0.001