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. 2007 Dec 13;104(51):20501–20506. doi: 10.1073/pnas.0710532105

Fig. 3.

Fig. 3.

Increasing knockdown of Bcr-Abl and of downstream targets STAT5 and CRKL in Ba/F3 p210-expressing cells transduced with single, double, and triple miRNA mimics. Ba/F3 Fluc-Neo cells (lane 1) expressing p210 IRES-EYFP (lanes 2–8) and grown in the presence of 10% IL-3-containing medium were infected with lentivirus expressing either miRNA Abl single 2 (lane 3), miRNA Abl double 6/2 (lane 4), miRNA Abl triple 6/2/1 (lane 5), miRNA scrambled (lane 6), shRNA Bcr-Abl junction (lane 7), or shRNA scrambled (lane 8). After sorting, cells were lysed for total protein in 2× sample buffer, and 5 × 104 cell equivalents were run per lane. Western blots were probed with 400 ng/ml anti-Abl (A), 1:1,000 anti-phosphotyrosine (B), 1:1,000 anti-STAT5 or 1:1,000 anti-phospho-STAT5 (C), 1:1,000 anti-CRKL or 1:1,000 anti-phospho-CRKL (D), 200 ng/ml anti-GFP (E), and 1:1,000 anti-ERK2 (F). Antibodies are described in Materials and Methods.