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. 2016 Aug 9;16:617. doi: 10.1186/s12885-016-2604-7

Fig. 5.

Fig. 5

Acetylated α-tubulin is regulated by HDAC6/ER-α interaction in RCC cell lines. a) HDAC 6, ER-α and acetylated α-tubulin protein expression were measured by western blot analysis in ccRCC tumors and adjacent non-tumor tissue. b) A representative ccRCC tumor showed HDAC 6 (in red) and ER-α (in green) localization in the cytoplasm. c-d) Representative immunofluorescent images of acetylated α-tubulin (in red) and HDAC 6 (in green) expression in parental and ER-α knockdown cell lines are shown. e) Knockdown of ER-α in MCF-7 and renal cell tumors as measured by Western blot analysis. f) Image J analysis measured immunofluorescence by calculating integrated density values of at least three representative fields per cell line. *p < 0.05 and **p < 0.01 indicates statistically significant difference of acetylated α-tubulin levels in ER-α knockdown cells as compared to the parental cell lines. The error bars represent standard errors from triplicate experiments and p-value was calculated using students t-test. g) C2H6 cells treatment with 10 μM hydroxy tamoxifen and/or 50nM panobinostat for 4 h. Representative immunofluorescence images of acetylated α-tubulin (in red), HDAC 6 (in green) and ER-α (in green) are shown. h) Image J analysis measured immunofluorescence by calculating integrated density values of at least three representative fields per cell line. *p < 0.01 indicates statistically different HDAC 6 levels in treated cell lines as compared to control cells. The error bars represent standard errors from biological triplicate experiments with technical replicates for each experiment. Scale bar indicates 50 μM distance and images are taken at 20X magnification