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. 2015 Aug 6;5:12917. doi: 10.1038/srep12917

Figure 4. Mutation of the deacetylase domain of HDAC6 abolishes its activities to deacetylate α-tubulin and cortactin and to induce ciliary disassembly.

Figure 4

(A) Western blot analysis of acetylated α-tubulin, α-tubulin, and GFP in RPE1 cells transfected with GFP vector or GFP-HDAC6 wild-type (WT), H216A, H611A, or H216/611A. (B) RPE1 cells were transfected with Flag-cortactin and GFP vector or GFP-HDAC6 WT, H216A, H611A, or H216/611A. The anti-Flag immunoprecipitate was then probed with acetylated lysine and Flag antibodies, and the cell lysate was probed with GFP antibody. (C–E) Immunofluorescence images (C), percentage of ciliated cells (D), and ciliary length (E) in RPE1 cells transfected with GFP vector or GFP-HDAC6 WT, H216A, H611A, or H216/611A, serum-starved for 24 hours, and stained with acetylated α-tubulin antibody and DAPI. Scale bar, 5 μm. *P < 0.05, ***P < 0.001; ns, not significant. Error bars indicate SEM.