Figure 5. IQDMA is a multi‐kinase inhibitor downregulating STAT5 nuclear levels.

• A
  Kinome screen with 10 μM IQDMA.
• B
  Subcellular fractions of SeAx cells treated with IQDMA for 24 h and immunoblotted for pY‐STAT5 and total STAT5. The cells were collected 30 min after 1 ng/ml IL‐2 cytokine addition. α‐Tubulin and histone H3 were used as loading controls for cytoplasmic and nuclear fractions, respectively. The normalized levels of phospho‐ and total STAT5 in the nucleus and cytoplasm, quantified by densitometry, are shown below the respective blots. One representative out of two independent experiments is shown.
• C
  Venn diagram showing the number of proteins that are significantly up‐/downregulated as a result of IQDMA treatments at 1, 2.5, and 10 μM.
• D–F
  Scatter plot depicting the profile of SeAx cells treated with (D) 1 μM IQDMA, (E) 2.5 μM IQDMA, and (F) 10 μM IQDMA. Protein abundances were determined using TMT‐based quantification mass spectrometry. Significant changes were assessed by a modified t‐test as implemented in the limma package, with the negative log₁₀ P‐values on the x‐axis, and log₂ fold change shown on the y‐axis.
• G
  Heatmap of IC₅₀ values calculated from drug response analysis using CellTiter‐Blue or CellTiter‐Glo viability assays upon 48 h drug treatment. One representative of three independent experiments performed in triplicates is shown. Gray represents the presence of STAT3/5 gain.

Source data are available online for this figure.