Effects of HG-7-85-01 on growth of nonmutated and mutant BCR-ABL–expressing cells and BCR-ABL kinase activity. (A) Determination of IC₅₀ values for HG-7-85-01 against parental Ba/F3 cells, Ba/F3.p210 cells, and Ba/F3-T315I cells. (B) ABL immunoprecipitation and pTYR and ABL immunoblots, respectively, of whole cell lysates prepared from Ba/F3.p210 cells treated for 2 hours with vehicle or HG-7-85-01 (0.1, 1μM). (C-D) Effects of HG-7-85-01 on proliferation of imatinib- and nilotinib-resistant BCR-ABL point mutant-expressing cells; 2- to 2.5-day treatments of BCR-ABL point mutant-expressing Ba/F3 cells with HG-7-85-01 in the absence (C) or presence (D) of IL-3. HG-7-85-01 treatment of parental Ba/F3 cells in the presence of IL-3 is shown in panels A and B as a control. Error bars represent the SEM for proliferation studies performed in duplicate. Assays for Ba/F3 cells expressing M351T, F317L, and F486S were carried out for 2 days. Assays for Ba/F3 cells expressing all other BCR-ABL point mutants were carried out for 2.5 days.