BC120 is stable and resistant to BC200 targeted RNAi. (A) T-47D cells were transfected with a BC200 expression plasmid. Forty-eight hours posttransfection, cells were treated with Actinomycin D to arrest transcription. Cells were harvested at the indicated time points posttreatment with Actinomycin D and BC200, and BC120 expression was monitored by northern blot with a probe targeting BC200 nt 63–84. Each RNA was quantified by densitometry relative to the 0 h time point. Data represent individual data points from three replicate samples with connecting line through the mean. (B) Representative northern blot used to generate the data shown in (A). (C) MCF-7 cells were transfected with LNA GapmeRs targeting the 5′ and 3′ ends of the BC200 RNA as well as a 3′ targeting siRNA and the indicated controls. Forty-eight hours posttransfection, cells were harvested, and total RNA extracted to perform northern blot with a probe targeting BC200 33–52. (D) Northern blot of RNA coimmunoprecipitated with antibodies to the indicated BC200-binding proteins using a probe complimentary to BC200 nt 33–52.
