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. 2012 Mar 30;7(3):e33662. doi: 10.1371/journal.pone.0033662

Figure 5. Analysis of acetylation and tyrosine phosphorylation of endogenous cortactin in WT and HDAC6-deficient MEFs.

Figure 5

(A) Isotype control (Ctrl.) and 4F11 immunoprecipitates from cell lysates of WT and HDAC6-deficient MEFs (H) were blotted first with acetyl-cortactin Ab (in green) and second with the 4F11 cortactin MoAb (in red). The merge of both images is shown. After gentle stripping to remove the acetyl signal, the membrane was blotted with pY466 Ab and 4F11 MoAb. Quantification and statistical analysis of three independent 4F11 immunoprecipitates and the ratio of acetyl:pY466 cortactin signals are shown. a.u.: arbitrary units. *, p<0.05. (B) Immunoprecipitates obtained with acetyl-cortactin Ab were blotted with pY466 Ab and 4F11. The phosphorylation signal did not coincide with acetylated cortactin. (C) Blotting of WT and HDAC6-deficient cell lysates with HDAC6 Ab is shown as a control of cell phenotype.