Molecular Characterization of Putative Chordoma Cell Lines
Brachyury expression in putative chordoma cell lines. (a) Immunoblot analysis of Brachyury of CCL4B, CM319, GB60, U-CH1, and U-CH2 cells reveals significant expression of the ~50 kDa Brachyury protein only in U-CH1 and U-CH2 cells. Molecular weight standard lane is denoted by M. (b) Immunofluorescent staining for Brachyury revealed nuclear localization of the protein in both U-CH1 and U-CH2. (c) Immunoblot analysis of cytokeratin protein expression in CCL4, CM319, GB60, U-CH1, and U-CH2. (d) Immunoblot analysis of PTEN protein expression in MRC-5 primary human fibroblasts, U-CH1 and U-CH2. (e) Immunoblot analysis of activation of Akt in normal human fibroblasts (growing, serum starved for 48 hours then with, or without, 10% added serum for 45 minutes), U-CH1 and U-CH2. Phospho-Akt- (Serine 473-) specific antibodies show Akt activation while similar protein loading was confirmed by staining for total Akt protein levels. For A, C, and E similar protein loading was confirmed by staining for β-actin.