Fig. 3

Thrombopoietin (TPO)-modulated intracellular Ca²⁺ flux in CALR mutated cells. (A) Schematic illustration of the experimental design of the study describing the TPO stimulation regimen. (B) Ca²⁺ influx in Fura-2 AM loaded 32D-CALR-WT/MPL, 32D-CALR-ins5/MPL and 32D-CALR-del52/MPL cells under starvation. After 3 h of IL3/FCS starvation, cells were stimulated with TPO (100 ng/ml) in Ca²⁺-free Ringer solution followed by addition of 2 mM extracellular Ca²⁺ (left panel). Bar graphs show Ca²⁺ flux response normalized to baseline Ca²⁺ levels (F340/380) as maximum peak and area under the curve (AUC) of store depletion (from 109 to 594 s) and SOCE (from 595 to 1080 s) (right panel). Data represents mean ± SEM from 5 independent experiments. Statistical analysis by Mann–Whitney-U-test. * p ≤ 0.05 (C) Western blot analysis of target proteins after 15 min of TPO (10 ng/ml) stimulation following 3 h IL3/FCS starvation. Numbers represent values normalized to the respective total protein