Journal: Leukemia

Article Title: A point mutation of zebrafish c-cbl gene in the ring finger domain produces a phenotype mimicking human myeloproliferative disease

doi: 10.1038/leu.2015.154

Figure Lengend Snippet: Increase in HSPCs was dependent on the Flt3 pathway. (a–d) flt3 MO injection attenuated the phenotype of LDD731. (e and f) c-myb staining of zebrafish embryos after treatment with 500 nM Lestaurtinib. (g and h) c-myb staining of embryos after treatment with 100 nM PKC412. (i) CBL mutation decreased FLT3 ubiquitination levels and upregulated mature FLT3. 293T cells were transfected with FLT3 and/or c-CBL constructs as indicated. After lysis, FLT3 was immunoprecipitated, blotted and probed with anti-FLT3 and anti-FK2. (j) Inhibitors were used to treat 293T cells. The cells transfected with FLT3 and c-CBL were treated with or without PKC412 or Lestaurtinib for 24 h. After lysis, protein samples were blotted and probed with FLT3, c-CBL and pFLT3 antibodies.

Article Snippet: FLT3 (fms-like tyrosine kinase 3) inhibitor PKC412 (ref. 22 ) (sc-200691) and Lestaurtinib 23 (sc-218657) with stock concentrations of 10 mM, were purchased from Santa Cruz Biotechnology and stored at − 80 °C.

Techniques: Injection, Staining, Mutagenesis, Ubiquitin Proteomics, Transfection, Construct, Lysis, Immunoprecipitation

Journal: bioRxiv

Article Title: Organelle proteomics reveals novel metabolic vulnerabilities in FLT3-ITD cells

doi: 10.64898/2026.01.19.700272

Figure Lengend Snippet: (A) Schematic overview of the organelle proteomics pipeline. FLT3 ITD-JMD (TKI-sensitive) and FLT3 ITD-TKD (TKI-resistant) Ba/F3 cells were left untreated or treated with 100 nM midostaurin for 24 h. Sequential extraction with six buffers generated six subcellular fractions, which were analyzed by MS-based proteomics in biological quadruplicates. Peptides were subjected to global proteomic profiling and phosphopeptide enrichment. (B) Intensity profiles of organelle-specific marker proteins across the six fractions. (C) Hierarchical clustering of protein profiles (scaled intensities) across fractions for untreated and midostaurin-treated FLT3-ITD cells. Statistically significant enriched gene ontology terms in each cluster are indicated.

Article Snippet: Midostaurin (Selleck chemical, S8064) was used at 100 nM for 24 hours.

Techniques: Extraction, Generated, Phospho-proteomics, Marker

Journal: bioRxiv

Article Title: Organelle proteomics reveals novel metabolic vulnerabilities in FLT3-ITD cells

doi: 10.64898/2026.01.19.700272

Figure Lengend Snippet: (A) Schematic representation of the bioinformatic strategy used to assign a cell compartment to proteins by using the SVM-algorithm. (B) Number of proteins assigned to cytosolic, nuclear, or organellar compartments in untreated and midostaurin treated FLT3-ITD cells. (C) Quantification of proteins changing their subcellular localization upon midostaurin treatment. (D) Sankey plot of protein translocation in midostaurin-treated FLT3 ITD-JMD . (E) Functional protein complexes redistributed between cytosol and nucleus in midostaurin-treated FLT3 ITD-JMD . (F) Sankey plot of protein translocation in midostaurin-treated FLT3 ITD-TKD cells. (G) Functional protein complexes redistributed between cytosol and nucleus in midostaurin-treated FLT3 ITD-TKD cells.

Article Snippet: Midostaurin (Selleck chemical, S8064) was used at 100 nM for 24 hours.

Techniques: Translocation Assay, Functional Assay

Journal: bioRxiv

Article Title: Organelle proteomics reveals novel metabolic vulnerabilities in FLT3-ITD cells

doi: 10.64898/2026.01.19.700272

Figure Lengend Snippet: Bar plots reporting the -log2 (p-value) of the GO-Term enrichment analysis of translocating proteins upon midostaurin treatment in sensitive and resistant FLT3-ITD cells. (C) Schematic representation of the spatial remodelling of key autophagic proteins upon midostaurin treatment in sensitive and resistant FLT3-ITD cells. (D) Heatmap reporting the intensity of the phosphorylation level of translocating autophagic proteins after midostaurin treatment in FLT3 ITD-JMD and FLT3 ITD-TKD . (E) Western blotting of LC3-II and p62 protein levels in FLT3 ITD-JMD and FLT3 ITD-TKD . Cells were treated with midostaurin with or without bafilomycin A1 for 24 hours. (F) Bar plot reporting the percentage of viable cells measured by MTT assay. FLT3 ITD-JMD and FLT3 ITD-TKD were treated with 100nm of midostaurin treatment for 24h either alone or combined with 3 hours of starvation.

Article Snippet: Midostaurin (Selleck chemical, S8064) was used at 100 nM for 24 hours.

Techniques: Phospho-proteomics, Western Blot, MTT Assay

Journal: bioRxiv

Article Title: Organelle proteomics reveals novel metabolic vulnerabilities in FLT3-ITD cells

doi: 10.64898/2026.01.19.700272

Figure Lengend Snippet: (A) Bar plot reporting the ECAR based measurements of glycolysis and glycolytic capacity in FLT3 ITD-JMD and FLT3 ITD-TKD . Cells were treated with 100nm midostaurin for 24 hours and subjected to a glycolysis stress test. ( B ) OCR-derived assessments of basal respiration and ATP production in FLT3 ITD-JMD and FLT3 ITD-TKD . Cells were treated with 100nm midostaurin for 24 hours and subjected to a mitochondrial stress test. (C) Representative electron microscopy images of steady-state FLT3 ITD-JMD and FLT3 ITD-TKD cells (m=mitochondria). (D) Bar plots reporting the number of cristae, the total cristae area and total mitochondria area in steady-state FLT3 ITD-JMD and FLT3 ITD-TKD cells. (E) Intensity profiles of the 168 proteins annotated in the MitoCarta database as mitochondrial proteins across the six fractions in each experimental condition. Colored boxes indicate the enriched GO-Term biological processes and cell compartments. (F) Heatmaps reporting the protein levels (scaled intensities) of the OXPHOS related proteins across the 6 fractions in each experimental condition.

Article Snippet: Midostaurin (Selleck chemical, S8064) was used at 100 nM for 24 hours.

Techniques: Derivative Assay, Electron Microscopy

Journal: bioRxiv

Article Title: Organelle proteomics reveals novel metabolic vulnerabilities in FLT3-ITD cells

doi: 10.64898/2026.01.19.700272

Figure Lengend Snippet: (A) Heatmap reporting the number of translocating proteins involved in carbohydrate, amino acids, peroxisome and lipid metabolism in FLT3 ITD-JMD and FLT3 ITD-TKD . (B) Bar plot reporting the ΔOCR of basal respiration after the inhibition of glucose usage (UK5099), glutamine usage (BPTES) and lipid usage (ETOMOXIR) for OXPHOS in FLT3 ITD-JMD and FLT3 ITD-TKD in absence and presence of 100nM midostaurin treatment for 24h. (C) Bar plot reporting the percentage of viable cells measured by MTT assay. FLT3 ITD-JMD and FLT3 ITD-TKD were treated with 100nm of midostaurin treatment for 24h either alone or combined with glucose, amino acid and lipid deprivation. (D) Representative immunofluorescence images of FLT3 localization before and after lipid deprivation in both permeabilized and non-permealized FLT3 ITD-JMD and FLT3 ITD-TKD . (E) Barplots showing the percentage of treatment-induced apoptosis (100 * (dead cells after treatment – death cells in control) / viable cells in control) in patient-derived blasts carrying FLT3-ITD in i) the JM domain, ii) both the JM and TK domain and iii) the TK domain upon the indicated treatments. Percentage of apoptotic cells was assessed by Annexin-V labeling.

Article Snippet: Midostaurin (Selleck chemical, S8064) was used at 100 nM for 24 hours.

Techniques: Inhibition, MTT Assay, Immunofluorescence, Control, Derivative Assay, Labeling

Journal: Cancer cell

Article Title: MAST1 drives cisplatin resistance in human cancers by rewiring cRaf independent MEK activation

doi: 10.1016/j.ccell.2018.06.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Midostaurin , LC Laboratories , Cat#P-7600; CAS: 120685–11-2.

Techniques: Recombinant, Negative Control, Viability Assay, Kinase Assay, Extraction, Ab Array, Activation Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription, shRNA, Sequencing, Plasmid Preparation, Software