Fig. 6: Bromodomain inhibitor iBET151 attenuates liver CXCL production and neutrophil infiltration in a murine alcohol feeding/LPS model.

a Schematic of alcohol feeding protocol. Mice were fed a 5% alcohol diet or a pair-fed control diet for 10 days. A subset of mice also received IP injection of the iBET151 compound. Alcohol-fed mice also received IP injection of LPS on day 11, 8 h before sacrifice (n = 12 pair-fed/Veh, n = 14 EtOH+LPS/Veh, n = 8 Pair-fed/iBET151, n = 6 EtOH+LPS/iBET151, same n for remaining panels of this figure). b qPCRs demonstrated CXCL chemokine and neutrophil marker Ly6g elevation with alcohol/LPS treatment. This response was attenuated by iBET151 injections. Expression levels were normalized to the average expression of pair-fed control mice. c IHC for MPO (neutrophil marker) is shown, with a number of neutrophils per low power field on the y-axis. The experiment was repeated with similar results. Scale bar = 100 μm. d Frozen section of mouse liver was stained with BODIPY 493/503 (green) and DAPI was used to stain nuclei (blue). The experiment was repeated with similar results. Scale bar = 100 μm. e No statistical difference was noted in serum ALT levels among groups. f Quantification of Western Blotting of cleaved caspase3 showed increased cleaved caspase 3 in livers of alcohol-fed/LPS mice and attenuated by iBET151 injection. For all analysis, two-way ANOVA was performed on normalized expression values for qPCRs, cell counts for IHC staining, normalized quantification of BODIPY 493/503 staining, ALT values, or quantification of WB bands with post hoc Tukey’s multiple comparison correction. Data represented as mean ± SD.