Figure 3.

Histone hypoacetylation inhibits LAT transcription in lung T cells after OVA exposure. A) The mRNA expression of LAT was determined by RT-PCR in lung T cells isolated from OVA-immunized rats. Non-immunized rats were used as control. The mRNA expression of LAT was normalized to β-actin and shown as the relative content of LAT mRNA. B) and C) Immunoblot analysis of LAT expression in lung T cells isolated from immunized or non-immunized rats. Data is shown as LAT/GAPDH ratio. D) and E) Western blots analysis of the protein level of HDAC1, H3Ac, H4Ac and H3K9me2 in lung T cells isolated from immunized rats and non-immunized ones. GAPDH was used as a loading control. Data is shown as relative percentage. F) and G) ChIP assays of histone acetylation and methylation status on LAT promoter in lung T cells isolated from immunized rats and non-immunized ones. The data represents agarose gel electrophoretic separation of amplicons obtained after PCR (a representative experiment from a series of three is shown). Input DNA served as positive internal control for sample integrity. Data is shown as mean ± SD of triplicate samples. *p < 0.05; **p < 0.01; P-value were calculated from two or three independent experiments with consistent results.