Journal: Signal Transduction and Targeted Therapy

Article Title: BMX inhibition overcomes small cell lung cancer chemoresistance by stabilizing E2F1 via ERK1/2-Cyclin D1/CDK4/6 axis

doi: 10.1038/s41392-026-02644-1

Figure Lengend Snippet: Characterization of IHMT-15137 as a potent and selective BMX inhibitor. a Chemical structure of IHMT-15137 and its reversible analog IHMT-15138. b Determination of the IC 50 values of IHMT-15137, IHMT-15138 and ibrutinib against the BMX protein via the ADP-Glo kinase assay. c Anti-proliferative activity of IHMT-15137, IHMT-15138 and ibrutinib against TEL-BaF3-BMX cells determined by the CTG assay. d CETSA analysis of the effects of IHMT-15137 on BMX protein stability in H446DDPR and H69AR cell lysates: temperature- and dose-dependent stabilization of BMX by IHMT-15137. The right panels show the results of the quantitative analysis of the BMX protein levels from the Western blots. e DARTS assay confirmation of the direct binding of IHMT-15137 to BMX by assessing the stability of BMX against pronase-mediated proteolysis in H446DDPR and H69AR cell lysates. f Inhibitory effects of IHMT-15137 and IHMT-15138 on BMX phosphorylation (Tyr566) in TEL-BaF3-BMX cells after 4 h of treatment, as determined by Western blotting. g Inhibitory effects of 12 h of IHMT-15137 treatment on BMX phosphorylation (Tyr566) and its downstream signaling pathways in H446DDPR and H69AR cells. h Molecular docking analysis of the irreversible binding mode of IHMT-15137 with BMX (PDB ID: 3SXR). i Effect of IHMT-15137 on Biotin-PEG2-C2-iodoacetamide labeling of BMX via a competitive biotin-iodoacetamide assay. Data were presented as mean ± SEM ( n = 3)

Article Snippet: Subsequently, the samples were incubated with 8 mM Biotin‐PEG2‐C2‐iodoacetamide (TargetMol, Shanghai, China; Cat. No. T17565 ) for 4 h at room temperature in the dark.

Techniques: Kinase Assay, Activity Assay, CTG Assay, Western Blot, Binding Assay, Phospho-proteomics, Protein-Protein interactions, Labeling