Figure 1.

ST2825 inhibits MYD88 oligomerization in L265P-expressing cells. (A) HEK293T cells were transfected with WT mCitrine TIR or an mCitrine-TIR L265P mutant. After 24 h of transfection, the cells were incubated for a further 24 h with DMSO or 10 µM ST2825. Following incubation, the cells were fixed and visualized under a confocal microscope at ×400 magnification. The representative images are provided from ≥3 independent experiments. (B, C) The cell lysate and pellet fractions were separated by centrifugation and analyzed using SDS-PAGE followed by anti-MyD88 and anti-β-actin immunoblotting. Relative MYD88 and β-actin expression levels were quantified and plotted as the β-actin-normalized ratios of the MYD88 pellet vs. lysate. Comparison of the pellet vs. lysate ratio in (B) OCI-LY10 and (C) TMD8 cells following treatment with DMSO or ST2825. Data are representative of three independent experiments expressed as the mean ± standard deviation, and a single representative immunoblot. **P<0.01. MYD88, myeloid differentiation primary response gene 88; TIR, Toll/interleukin-1 receptor; WT, wild-type; WCL, whole cell lysate; IB, immunoblot.