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. 2014 Mar 6;9(3):e90850. doi: 10.1371/journal.pone.0090850

Figure 9. Effects of berberine (BBR) to inhibit the degradation of RB1 mRNA with poly(A) (RB1-pro-RB1) or without poly(A) (RB1-pro-RB1-Δpoly A-).

Figure 9

(A) Cartoon of the plasmid models used in this study. Polyadenylation signals were showed in red. (B) Expressions of GFP in cells with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids detected using Western blot. (C) Expressions of β-actin and RB1 mRNA using random primer and oligo d(T) in the cells with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids detected using RT PCR. (D) Fluorescence intensity in cells with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids detected using flow cytometry. (E) RB1 mRNA expressions using oligo d(T) as the primer in cells with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids detected using real time PCR. (F) RB1 mRNA level in PC12 cells in normal conditions with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids administrated with or without BBR. (J) RB1 mRNA level in PC12 cells in oxygen and glucose deprivation (OGD) conditions with transfection of RB1-promoter-RB1-GFP with and without poly(A) plasmids administrated with or without BBR. (H) The fluorescence spectrum of BBR associating with poly(A) (40 µM). BBR was 0.5 µg/ml. Data are presented as mean ± S.D. from three independent experiments (n = 3). **P<0.01.