Briefly, the intensity of the bands were measured by Image J software, Bethesda, MD, USA and lamin Vandetanib cost A, a nuclear protein, was used for Inhibitors,Modulators,Libraries loading controls. Statistical analysis All results Inhibitors,Modulators,Libraries are expressed as means SE. Statistical signifi cance of differences between two groups was determined by the Mann Whitney U test. Other analysis was per formed to compare differences among groups by one way analysis of variance followed by the Bonnferoni correc tion. A P value of less than 0. 05 was considered statisti cally significant. Results Measurement of HDAC activity in RA, OA and normal synovial tissues Total nuclear HDAC activity in samples of synovial tissue from RA patients was 0. 96 0. 08 uM of the HDAC stan dard. This level of activity was significantly higher than those from OA and from normal controls.

Measurement of HAT activity in RA, OA and normal Inhibitors,Modulators,Libraries synovial tissues Total nuclear HAT activity was measured in synovial tis sues from normal Inhibitors,Modulators,Libraries controls, from OA and from RA. There was no Inhibitors,Modulators,Libraries significant difference between normal controls, OA and RA synovial tissues. We examined the ratio of HDAC to HAT activity on the same patients, but failed to show the significant difference in the ratio of HDAC to HAT activity between OA and RA groups. This might be partly due to small data set, but, at least, HDAC HAT was not shifted toward histone hyperacetylation. To establish the relationship between nuclear HDAC activity and cytoplasmic TNF levels directly, we mea sured TNF of the cytoplasmic fraction which was obtained by preparing nuclear extracts of OA and RA synovial tissues.

The amount of cytoplas mic TNF tended to correlate with nuclear HDAC activ ity. Class I, class II HDACs and TNF mRNA expressions in total synovial tissue To investigate the expression profiles of class I and class II HDACs in inhibitor Trichostatin A RA, OA and normal controls synovial tissues, mRNA levels of HDAC1 to 8 were evaluated by quantitative real time PCR. RA synovial tis sues expressed high levels of HDAC1 compared to OA, and normal controls. HDAC4 mRNA levels were significantly higher in normal controls than in RA. TNF mRNA expression was measured in RA synovial tissue. The data showed significant posi tive correlation between TNF and HDAC1 mRNA in RA synovial tissue . Nuclear expression of class I HDACs in synovial tissue We performed Western blotting for nuclear class I HDACs in synovial tissue. The level of nuclear HDAC1 protein expression was higher in RA synovial tissue com pared with OA synovial tissue. Western blots were quantified by Image J software. The level of expression of HDAC1 by normalizing to the band density of nuclear membrane protein lamin A was significantly higher in RA than OA synovial tissue .