Fig. 4
BBR promotes UHRF1 protein degradation and reactivates several TSGs. a BBR reduced the expression of UHRF1 and DNMT1 proteins. UHRF1 in RPMI-8266, MM.1S, and U266 cells were treated with 25 μM BBR for 0, 4, 8, 12, 24, and 48 h. Cell lysates were harvested and subjected to western blotting with anti-UHRF1, anti-DNMT1, and anti-GAPDH antibodies. b, c Screening the effective UHRF1-siRNA in MM cells. UHRF1-siRNA #2 was confirmed as the effective UHRF1-siRNA through qRT-PCR and western blotting after transfection with 100 nM UHRF1-siRNAs for 48 h. The data were presented as the mean ± SD obtained from three independent experiments. d, e The effect of BBR on the expression of UHRF1 at 48 h. The mRNA and protein expression of UHRF1 in MM cells was determined through qRT-PCR and western blotting after treatment with or without 25 μM BBR for 48 h. The data were presented as the mean ± SD obtained from three independent experiments. f, g The effect of BBR on the expression of p16INK4A, p53, and p73. The mRNA and protein expression of p16INK4A, p53, and p73 in MM cells were determined through qRT-PCR and western blotting after treatment with or without 25 μM BBR for 48 h. The data were presented as the mean ± SD obtained from three independent experiments