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. 2001 Nov;21(21):7416–7428. doi: 10.1128/MCB.21.21.7416-7428.2001

FIG. 4.

FIG. 4

Effects of E12 on endogenous p21 expression and BrdU incorporation in MG63 cells. (A) Activation of endogenous p21 by E12 overexpression. MG63 cells were transfected with pCMV-E12 and maintained in DMEM containing 10% FBS for 72 h. The cells were fixed and stained for E12 (rhodamine; a), p21 (fluorescein; b), and DNA (DAPI; c). DAPI was used for counterstaining to visualize the nucleus. Arrowheads, cells transfected with E12. (B) Effects of E12 on cell proliferation. MG63 cells were treated by the same procedure as for panel A. BrdU was added 1 h before fixation. Arrowheads, positions corresponding to cells. The cells were stained for E12 (rhodamine; a), BrdU (fluorescein; b), and DNA (DAPI; c). (C) Impairment of E12-dependent p21 induction by the p21 antisense plasmid. MG63 cells were transfected with pCMV-E12 and an antisense p21 construct (pCMV-p21as) and maintained in DMEM containing 10% FBS for 72 h. The cells were fixed and stained for E12 (rhodamine; a), p21 (fluorescein; b), and DNA (DAPI; c). Arrowheads, cells expressing E12. (D) Impairment of E12-mediated cell cycle attenuation by the p21 antisense plasmid. MG63 cells were treated by the same procedure as for panel C. BrdU was added 1 h before fixation. Arrowheads, positions corresponding to cells. The cells were stained for E12 (rhodamine; a), BrdU (fluorescein; b), and DNA (DAPI; c). Scale bar, 20 μm.