Fig. 3

HDAC 1 and HDAC 6 increase MMP-2/9 activity and decrease acetylated α-tubulin levels, respectively. a) C2 and 786–0 parental and knocked down HDAC 1 were analyzed for MMP activity by gelatin zymography assay. MMP activity was measured both in cell lysates as well as cell supernatants. b) Renal tumor cell lines were analyzed for HDAC 6 and acetylated α-tubulin expression by immunofluorescence. The staining in red indicates acetylated α-tubulin and the staining in green indicates HDAC 6 expression. Scale bar indicates 50 μM distance and images are taken at 20X magnification. c) Image J analysis measured immunofluorescence by calculating integrated density values of at least three representative fields per cell line. *p < 0.05 indicates statistically significant difference in acetylated α-tubulin levels as compared to C2 cells. The error bars represent standard errors from triplicate experiments and p-value was calculated using students t-test