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Roles of ALDH in T-ALL
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Figure 3. ALDH1A2 supports leukemia cell survival in T-cell acute lym- phoblastic leukemia cells. (A) A scheme of the doxycycline (Dox)-inducible sgRNA-guided dCas9-KRAB system (left). Two independent sgRNA targeting the ALDH1A2 regulatory element (sgRNA #1 and #2) were induced by Dox treatment. Control cells were treated with the vehicle (dimethylsulfoxide, DMSO). (B) Western blot analysis showing protein expression of ALDH1A2, PARP, caspase 3 and α-tubulin in Jurkat cells at 48-96 hours after Dox treat- ment. (C) The apoptotic cell population (%) was determined based on the number of annexin V-positive and propidium iodide-negative cells in multiple T-cell acute lymphoblastic leukemia (T-ALL) cell lines on day 5 after the trans- duction of lentiviral shRNA targeting ALDH1A2 or GFP (negative control). Error bars represent the standard deviation (SD) for technical replicates. *P<0.05, **P<0.01 using the two-tailed Student t-test. (D) Cell viability was measured by a CellTiter Glo assay on day 4 after shRNA knockdown of ALDH1A2 in Jurkat cells. Error bars represent the SD for technical replicates. ***P<0.001 using the two-tailed Student t-test. (E) Cell viability was measured by a CellTiter Glo assay on day 4 after the induction of ALDH1A2 expression by Dox treatment in DND-41 cells. Error bars represent the SD for technical repli- cates. *P<0.05 using the two-tailed Student t-test. (F) Cell viability was meas- ured by CellTiter Glo assays on day 7 after treatment with WIN 18,446 at dif- ferent doses in two TAL1/ALDH1A2-positive (Jurkat and RPMI-8402) and two TAL1/ALDH1A2-negative (KOPT-K1 and DND-41) T-ALL cell lines. Nonlinear regression fit curves are shown. Error bars represent the standard error of the mean for technical replicates.
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