Fig. 6: ROCK1/2-inhibition alters the gene expression profile in vivo and reduces NF-κB and cofilin phosporylation in DCs and in macrophages in vitro.

a The heatmap shows the expression of downregulated genes related to PI3K signaling in CD45⁺CD11b⁺ cells that were FACS-isolated from mice on day 14 after allo-HCT after 10 days of ROCK1/2-inhibitor or vehicle treatment. Color scale represents the row-wise z-score intensity. * Indicates significant changes in gene expression. b The heatmap shows the expression of genes involved in Tnf signaling of CD45⁺CD11b⁺ cells isolated as indicated in (a). Color scale represents the row-wise z-score intensity. * Indicates significant changes in gene expression. Red arrow indicates NF-κB1 gene. c–f The western blots show total and phosphorylated NF-κB of BM-derived DCs or macrophages incubated with LPS for two hours and thereafter with ROCK1/2-inhibitor for two hours. Samples derived from the same experiment and gels/blots were processed in parallel. Loading controls ran on the same blot. When applicable, data is normalized to control group, the experiments were performed three times, each point represents a mouse, values of the experimental group were normalized to control group. The plots show mean +/− SD and the P-value was calculated using a two-tailed paired student t-test. Uncropped western blots are shown in Source Data. c The bar graph shows pNF-κB/total NF-κB in BM-derived DCs. d Representative western blot image of pNF-κB and total NF-κB in BM derived DCs. e The bar graph shows pNF-κB/total NF-κB in BM-derived macrophages. f Representative western blot image of pNF-κB and total NF-κB in BM-derived macrophages. g–j The western blots show total and phosphorylated Cofilin of BM-derived DCs or macrophages incubated with LPS for two hours and thereafter with ROCK1/2-inhibitor for two hours. Samples derived from the same experiment and gels/blots were processed in parallel. Loading controls ran on the same blot. When applicable, data are normalized to control group, the experiments were performed three times, each point represents a mouse, values of the experimental group were normalized to control group. The plots show mean +/− SD and the P-value was calculated using a two-tailed paired student t-test. Uncropped western blots are shown in Source Data. g The bar graph shows pCofilin/total Cofilin in BM-derived DCs. h Representative western blot image of pCofilin and total Cofilin in BM-derived DCs. i The bar graph shows pCofilin/total Cofilin in BM-derived macrophages. j Representative western blot image of pCofilin and total Cofilin in BM-derived macrophages.