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. 2014 Oct 6;5:5059. doi: 10.1038/ncomms6059

Figure 4. ABRO1 increases the protein stability of p53.

Figure 4

(a). HCT116 p53+/+, HepG2 and A549 cells were transfected with increasing concentrations of plasmid encoding Flag–ABRO1. After 36 h, proteins were extracted and subjected to western blotting. (b) HCT116 p53+/+ cells were transfected with different ABRO1 siRNAs and siNC. After 36 h, proteins were extracted and subjected to western blotting. (c) HCT116 p53+/+ cells were co-transfected with plasmids encoding Flag-ABRO1 and Myc-MDM2. Proteins were extracted and subjected to western blotting. (d) HCT116 p53+/+ cells were treated with cycloheximide (CHX) (1 μg μl−1) at the indicated times after transfection with pCMVFlag-ABRO1. Proteins were extracted and subjected to western blotting. (e) HCT116 p53+/+ cells were treated with CHX (1 μg μl−1) at the indicated times after transfection with siABRO1-2 or siNC. Proteins were extracted and subjected to western blotting. (f) HCT116 p53+/+ cells were co-transfected with the indicated constructs, and reporter activity was determined as described in Experimental Procedures. Data were generated from three independent experiments, each performed in triplicate (The data shown are the mean±s.d.). Data were analysed using Student’s t-test.*P<0.05, **P<0.01.