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Fig. 1 | BMC Cell Biology

Fig. 1

From: Acetylation of C-terminal lysines modulates protein turnover and stability of Connexin-32

Fig. 1

HDAC6 inhibition reduces turnover and increases acetylation of Cx32. a TSA and (b) TubA increase expression level of Cx32. Western blots of N2a cell lysates treated with the TSA (concentrations as indicated) (a) or 20 μM TubA for 6 h (b) were probed with anti-Cx32, actin (a) or tubulin (b) was used as a loading control. c TubA treatment decreases turnover of Cx32 (bottom panel) compared to untreated samples (top panel). Protein turnover of 35-S-Cx32 was determined by phosphorimaging at 0, 1, 2, 4 h post-chase (shown). Blots were subsequently probed with anti-Myc as a loading control. d Relative levels of 35-S-Cx32 remaining after 1.5 h post-chase (in subsequent experiments) was quantified by densitometry of phosphorimages, normalized to Myc, and used to calculate half-life, as described (see Methods). (N = 4 independent experiments; student’s T-test* p < 0.05 as compared to DMSO control). e TubA increases expression levels of total Cx32 and acetylated Cx32. Cx32 immunoprecipitates of N2a cell lysates expressing Cx32-Myc/6xHis treated with TubA or vehicle alone were processed for Western blots analysis and probed with Cx32 and AcK antibodies

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