Research Article

Ubiquitin-Specific Protease 2 Modulates the Lipopolysaccharide-Elicited Expression of Proinflammatory Cytokines in Macrophage-like HL-60 Cells

Figure 3

USP2 variants modulate expression of proinflammatory cytokines. Cells were treated with PMA (30 nM; 1 d) and subsequently stimulated with LPS (2.5 μg/mL) or vehicle alone for (a–d) 4 h or (e) 12 h. Transcript levels of (a) USP2 splice variants and (d) cytokines were determined by qRT-PCR and are normalized according to HPRT-1 transcript levels. (b) Western blot analysis of USP2 protein in the USP2 variant-restored cells stimulated with LPS. Signal intensities of the bands were measured and normalized to the level of GAPDH. Representative images are shown. (c) Schematic summary of cytokine expression analysis in the USP2 variant-restored cells. Four biological replicates were used for this analysis. (e) The IL-6 concentration in the supernatant of USP2 variant-restored cells was determined by ELISA. Data are presented as means ± SD of (a, d) six, (b) three, or (e) eight samples. Mock, USP2KD cells with a mock vector; AR, USP2KD cells with a USP2A expression construct; BR, USP2KD cells with a USP2B expression construct. versus mock-transfected control cells; # versus USP2AR cells; § versus vehicle-treated cells.