Figure 7. Effect of KDM4B knockdown or ML324, a pharmacological inhibitor of KDM4, on TNF-α-induced leukocyte adhesion and transmigration.

(a) Cells transfected with 100 nM scrambled or 100 nM KDM4B siRNA were treated with Tris-HCl, pH 8.0 buffer or TNF-α for 24 h and then exposed to 10⁵ HL-60 cells at 37 °C for 1 h. (b) Cells pretreated with ML324 (0.2 μM) for 1 h were treated with TNF-α for 24 h and then exposed to10⁵ HL-60 cells at 37 °C for 1 h. Attached HL-60 cells were counted under an inverted microscope at 400× magnification (mean ± SEM; n = 4–5). Scale bars, 50 μm. (c) Quantitation of HL-60 cell transmigration assay. The bar graph indicates the number of migrating HL-60 cells (mean ± SEM; n = 3–4). (d) Neutrophil adherence to brain microvessels. The bar graph indicates the attached neutrophil number per mm² (mean ± SEM; n = 3). Scale bars, 20 μm.