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. 2010 Dec 20;31(4):686–699. doi: 10.1128/MCB.00019-10

FIG. 9.

FIG. 9.

Functional importance of Sox-binding sites in cotransfection assays. (A to C) AC8Luc was cotransfected with Sox expression plasmids in various cultures as indicated. Western analysis with anti-FLAG antibody (C) shows the relative expressions of L-Sox5/Sox6 and SOX9 in the transfected COS-7 samples. (D) Effect of point mutations on the synergistic activation of the long promoter by L-Sox5/Sox6 and SOX9 coexpressed at optimal (2.7:1) molar ratio in COS-7 cells and at a higher ratio in LDM and CEC cultures. The schematic indicates factor binding to the short promoter elements and to the upstream elements (Upe) (not drawn to scale). Thin and thick arrows depict the transcription efficiencies at early (E) and at late (L) stages of chondrogenesis. Luciferase activities are given as fold values relative to that for AC8Luc. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (compared with the reporter cotransfected with vectors [A to C] or between the cotransfected mutants and the similarly cotransfected wild-type AC8Luc [D]); #, P < 0.05; ##, P < 0.01; ###, P < 0.001 (compared with the SOX9-cotransfected reporters).