Fig. 2.

Efficient knockdown of multiple chimeric forms of cAbl using single, double, and triple miRNA mimics. Five micrograms of MSCV-IRES-EYFP expressing either p210 Bcr-Abl WT, p210 Bcr-Abl T315I (ref. 41), Tel-Abl (ref. 39), or p185 Bcr-Abl WT (ref. 61) were transfected alone (lane 1) or cotransfected with 5 μg of either miRNA scrambled (lane 2), miRNA Abl single 2 (lane 3), miRNA Abl double 6/2 (lane 4), or miRNA Abl triple 6/2/1 (lane 5) onto 293T cells. All miRNAs were in the pcDNA 6.2-GW/EmGFP vector. Forty-eight hours after transfection, cells were lysed in extraction buffer as described in Materials and Methods, and 5 μg of protein was run per lane. Blots were probed with 400 ng/ml anti-Abl and 1:1,000 anti-ERK2 as described in Materials and Methods.