Figure 6.

Depletion or knockdown of HDAC6 increases endogenous ERK1/2 acetylation. A, knockout of HDAC6 increases ERK1/2 acetylation. Left panels, anti-AcK antibody was used to immunoprecipitate acetylated ERK1/2 in HDAC6 wildtype or HDAC6 knockout MEFs. The immunoprecipitates were resolved on SDS-PAGE, and the anti-ERK1/2 Western blot analysis was performed. 0.5% of whole-cell lysate was used as the input. Western blot analyses were performed using anti-ERK1/2, anti-HDAC6, anti-acetyl–α-tubulin, or α-tubulin antibodies as indicated. Right panel, bar graph was used to show relative intensity of endogenous ERK1 and ERK2. Five independent experiments were performed. B, knockdown of HDAC6 increases ERK1/2 acetylation. The experiments were conducted the same as A except that A549 control and A549 HDAC6-KD pair were used to replace the wildtype and HDAC6KO MEFs pair. Four independent experiments were performed. Student's t tests were performed with *, p < 0.05; **, p < 0.01. Error bars, S.D. IB, immunoblot.