Figure 1 | Leukemia

Figure 1

From: Mutant JAK3 phosphoproteomic profiling predicts synergism between JAK3 inhibitors and MEK/BCL2 inhibitors for the treatment of T-cell acute lymphoblastic leukemia

Figure 1

Experimental workflow. Three independent sets of Ba/F3 cells expressing mutant human JAK3 (L857Q) were cultured and treated with 500 nM tofacitinib, ruxolitinib or dimethylsulfoxide (DMSO) for 90 min (biological and technical replicates). Cells were digested with Typ/Lys-C mix. Peptides from each of the nine samples were labeled using tandem mass tags (TMT-10plex, Thermo Fisher Scientific) and mixed 1:1. Phosphopeptides were isolated and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed using a Q-Exactive Plus hybrid quadrupole-Orbitrap MS system (Thermo Fisher Scientific). Database searching of all.raw files was performed using Proteome Discoverer 2.1 (Thermo Fisher Scientific). Mascot 2.2.3 and SEQUEST HT were used to search against the Swiss_Prot, Uniprot_mouse database.

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