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. 2014 Jan 2;9(1):e84460. doi: 10.1371/journal.pone.0084460

Figure 5. Necdin and Bmi1 relieve transcriptional suppression of the p16 and Cdk1 promoters.

Figure 5

(A, B) BrdU incorporation assay. HEK293A cells transfected with expression vectors for GFP, necdin, and Bmi1 were immunostained for GFP (green) and BrdU (red) (A). BrdU+ cells among 500 GFP-expressing cells were counted (n = 3) (B). Arrowheads point to GFP+/BrdU+ cells. Scale bar, 10 µm. (C, D): Promoter assay. Luciferase reporter vectors containing the Cdk1 (C) and p16 (D) promoters and expression vectors for E2F1, necdin, and Myc-tagged full-length Bmi1 (Bmi1) were co-transfected into HEK293A cells. Promoter activities were measured with a luminometer. Values in (B−D) represent the mean ± SEM, n = 3. *p<0.05, **p<0.01.