Functional and molecular characterization of Ba/F3 FLT3-ITD-R1-4 cells
. | IC50 . | . | FLT3 expression . | . | . | ||
|---|---|---|---|---|---|---|---|
| Cell line . | SU5614, μM . | AraC, μg/mL . | FACS, MCF . | WB, % of ITD native . | FLTc-TKD mutation . | ||
| ITD native | 0.2 | 0.06 | 14.1 | 100 | — | ||
| ITD-R1 | 4.5 | 0.03 | 50.9 | 725 | Y842H | ||
| ITD-R2 | 5.2 | 0.08 | 41.2 | 918 | Y842H | ||
| ITD-R3 | 2.1 | ND | 63.3 | 1016 | D835N | ||
| ITD-R4 | 1.3 | 0.04 | 54.4 | 1285 | D835N | ||
. | IC50 . | . | FLT3 expression . | . | . | ||
|---|---|---|---|---|---|---|---|
| Cell line . | SU5614, μM . | AraC, μg/mL . | FACS, MCF . | WB, % of ITD native . | FLTc-TKD mutation . | ||
| ITD native | 0.2 | 0.06 | 14.1 | 100 | — | ||
| ITD-R1 | 4.5 | 0.03 | 50.9 | 725 | Y842H | ||
| ITD-R2 | 5.2 | 0.08 | 41.2 | 918 | Y842H | ||
| ITD-R3 | 2.1 | ND | 63.3 | 1016 | D835N | ||
| ITD-R4 | 1.3 | 0.04 | 54.4 | 1285 | D835N | ||
The IC50 of SU5614 and Ara-C in ITD-R1-4 cells was determined as described in Figure 4. FLT3 expression was measured by FACS, and the mean channel fluorescence (MCF) was calculated. The expression of FLT3 (WB) was determined by Western blot analysis with the use of an anti-FLT3 antibody. The blot was stripped and reprobed with an anti–β-actin antibody. The results were quantified by densitometry according to the FLT3–β-actin ratio of parental cells that was set to 100%. FLT3-TKD mutation indicates the presence (Y842H and D835N) or absence (—) of point mutations in TKD. ND indicates not done.