The intermediate filament cytoskeleton in epithelial tissues is formed of keratin heterodimers. Keratins are highly post-translationally modified proteins, with tyrosine phopsphorylation, serine phosphorylation, and glycosylation amongst reported modifications. We and others have recently reported multiple acetylation sites on keratin 8 and we have previously shown that these sites are responsive to butyrate. In this study, we report the application of cellomic approaches to demonstrate differential responses of three lysine acetylations (lys 10, lys 471 and lys 482) to different short-chain fatty acids. The data imply no fixed hierarchy of acetylation on keratin 8, and furthermore imply different ranges of histone deacetylase (HDAC) inhibitory specificities for short chain fatty acids (SCFA). Furthermore we have used the functionality of the High Content Analysis (HCA) platform to show that the acetylation sites are differentially modified in cells undergoing mitosis. Taken together the data imply distinct roles for keratin acetylations in function