Figure 3.

Impact of tunicamycin or 2-deoxy-D-glucose on signal transduction, apoptosis, and proliferation of human FLT3-ITD-expressing MOLM13 cells. Human FLT3-ITD expressing MOLM13 cells were pre-treated for indicated combinations either with 50 nM tunicamycin (A) or 3 mM 2-deoxy-D-glucose (B) for 24 h after 75 nM 17-AAG was added to the wells for further 24 h. The protein lysates were subjected to immunoblotting to assess changes of indicated signalling proteins. For loading control, levels of GAPDH were detected. The corresponding apoptosis rates are shown after the cells were stained with Annexin V and subsequently analyzed by flow cytometry (upper right of A and B). For same conditions, a PrestoBlue viability assay was carried out and the dataset that is normalized to the control is illustrated (center right A and B). IC₅₀ values are indicated for 17-AAG dependence on different concentrations of pre-treatment with tunicamycin and 2-deoxy-D-glucose (lower right, A and B). Student’s t-test was used for calculating the statistical significance (**** p < 0.0001, *** p < 0.001, ** p < 0.01, ns not significant). The data for the MTS assays are shown as relative absorption compared to untreated controls. The histograms provide representative data of triplicates that were reproduced in three independent experiments. (mean values and ± SD). The replicates are shown in the Supplement Table S3.