Figure 1: MS-275 affects inflammatory gene expression and histone acetylation in precision-cut lung slices. | Scientific Reports

Figure 1: MS-275 affects inflammatory gene expression and histone acetylation in precision-cut lung slices.

From: HDAC1-3 inhibitor MS-275 enhances IL10 expression in RAW264.7 macrophages and reduces cigarette smoke-induced airway inflammation in mice

Figure 1

Schematic representation of the experimental setup and timeline (A). Mouse precision-cut lung slices (PCLS) were subjected to MS-275 (1 μM or 10 μM) or SAHA (0.41 μM) for 20 h and stimulated with LPS/IFNγ the last 4 h of the experiment, and lysed. Gene expression was studied by RT-qPCR and expressed as fold change compared to control (LPS/IFNγ-treated) (B). Data are presented as mean ± SD of 5–11 independent experiments. *p < 0.05, **p < 0.01; ***p < 0.001 compared to vehicle (LPS/IFNγ and inhibitor solvent-treated). Under the same conditions, PCLS histones were extracted, resolved by SDS-PAGE, and effects of MS-275 on total histone acetylation were assessed by Western blot (dotted line indicates crop within the same blot; full length blot is presented in Figure S2) (C) and quantified by densitometry (D). Data are presented as mean ± SD of 2–3 independent experiments. *p < 0.05 compared to vehicle (LPS/IFNγ and inhibitor solvent-treated). Histones H3 and H4 were also excised from the gel and subjected to LC-MS/MS analysis (E). MS-275 increased acetylation on one peptide from histone H3 (res. peptide 18–26: KQLATKAAR) and one peptide from histone H4 (res. peptide 4–17: GKGGKGLGKGGAKR). Other peptides were not affected. Data are presented as mean ± SD of 3 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 compared to vehicle (LPS/IFNγ and inhibitor solvent-treated).

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