Fig 1: Inhibition of H4K12ac with NU9056 modulates cytokine and chemokine production. After 5–7 days of differentiation, MDDCs were treated with NU9056 50 nM, 0.2% EtOH or both for 5 days. Supernatants collected were analyzed for 48 inflammatory cytokines and chemokines. Data is from 3 blots for untreated control, 2 blots each for treatment with 0.2% EtOH alone, treatment with NU9056 alone, and treatment with 0.2% EtOH plus NU9056. Cytokines chosen to be presented in the graph were selected on the basis of fold change (2 folds or more) compared to control. Each cytokine is detected in duplicates within each blot. Statistical differences were calculated using student’s t-test when individually compared to untreated control and indicated with a * (compared to control) and # (between treatments) for significant p value. Two-way ANOVA was carried out (p = 0.027). Panel a shows optical density values for cytokines IL-15, RANTES, TGF-ß1 and TNF-a. Panel b shows optical density values for MCP2. In panel c, MCP-2 ELISA was carried out from cell culture medium at least three times in duplicates. Values detected in pg/mL are as follows, untreated control (322.4 ± 25.04), NU9056 50 nM (317.5 ± 14.3), 0.2% EtOH (401.7 ± 28.2) and 0.2% EtOH + NU9056 50 nM (321.6 ± 13.5, p = 0.03). Panels d–g show intracellular staining to measure MCP-2 levels in untreated or treated MDDCs. Data represented is from at least 3 experiments. Panel d shows % gated MCP-2 positive cells for untreated control (52.3% ± 2.3), NU9056 50 nM (45.8% ± 1.07), 0.2% EtOH (85.8% ± 5.2, p = 0.002) and 0.2% EtOH + NU9056 50 nM (29.6% ± 1.18, p = 0.0001). Panel e shows MFI values for MCP-2-FITC for untreated control (2510 ± 29), NU9056 50 nM (2151 ± 43), 0.2% EtOH (4260 ± 603, p = 0.04) and 0.2% EtOH + NU9056 50 nM (1459 ± 297, p = 0.01). Panel f shows an overlay histogram of MCP-2-FITC for all the samples as shown in legend. Panel g shows representative single cell images.