Figure 1. UNG siRNA inhibits expression of the UNG gene in human prostate cancer cells.

(A) Four individual siRNAs (siUNG) against the UNG gene were combined into one pool, and the human prostate cancer cell lines LNCaP, DU145 and PC3 were transfected with various amounts of the pooled siUNG as indicated. The mismatch control siMM (200 nM) was used as a negative control. Twenty-four hours after transfection, total cell extracts were isolated from controls and siUNG-transfected LNCaP, DU145 and PC3 cells, and the immunoblot analysis was performed using UNG and GAPDH antibodies. GAPDH was utilized as a loading control. The number under each band is expressed as a percentage of Silentfect control, normalized by the corresponding GAPDH level.

(B) Human prostate cancer cell lines LNCaP, DU145 and PC3 were transfected with siUNG (100 nM) and the cells were collected at different time points after transfection as indicated and immunoblot analysis for UNG and GAPDH was performed. The number under each band is expressed as a percentage of Silentfect control, normalized by the corresponding GAPDH level.

(C) Human prostate cancer cell lines LNCaP, DU145 and PC3 were transfected with siUNG (10–200 nM) or siMM (200 nM) as in A. Twenty-four hours after transfection, cells were collected, and the mRNA levels of UNG and GAPDH were examined by RT-PCR. Note that UNG represents the expression of both UNG1 and UNG2. The number under each band is expressed as a percentage of Silentfect control, normalized by the corresponding GAPDH level..

(D) Human prostate cancer cell lines LNCaP, DU145 and PC3were transfected with siUNG (100 nM) and the cells were collected at different time points after transfection as indicated and RT-PCR analysis for UNG and GAPDH was performed. The number under each band is expressed as a percentage of Silentfect control, normalized by the corresponding GAPDH level.