Figure 3.

Salarin C inhibits Culture Repopulation Ability of CML cell lines and rescues BCR/Abl signaling in CML cell lines incubated in low oxygen. (A) K562 or KCL22 cells were treated or not (control) at time 0 with a single dose of salarin C at the indicated concentration (μM) and incubated at 0.1% oxygen (LC1). On day 7 of LC1, cells were transferred to secondary cultures (LC2) established in the absence of salarin C and incubated in normoxia. Trypan blue-negative cells were counted at the indicated timepoints of LC2; the graphs represent average ± SEM of data from 3 independent experiments; significant differences are indicated (Student's t test for independent samples; *: p< 0.05). (B, C) Cells were treated or not at time 0 with the indicated concentrations of salarin C and incubated at 0.1% oxygen for the indicated times. Cells were lysed in Laemmli buffer and lysates subjected to immuno-blotting with antibodies for the indicated proteins. Due to the marked MW difference, anti-phospho-Abl antibody made it possible to detect phospho-BCR/Abl. Anti-GAPDH antibody was used to verify equalization of protein loading. One representative experiment out of 3 is shown.