Fig. 5
From: Loss of Kat2b impairs intraflagellar transport and the Hedgehog signaling pathway in primary cilia
The acetyltransferase domain of Kat2b in the cytosol is significant for localization of IFT components at primary cilia. (a) Graphical view of domains of Kat2b and designed mutant Kat2b. (b) Immunocytochemistry images confirmed the localization of the designed mutant Kat2b construct. DsRed tagged Kat2b constructs were investigated vector transfected cell and nuclear localization signal domain deleted Kat2b positioned only cytosol. (c) Immunoblotting analysis verified cytosolic and nucleic distribution under transfection with Kat2b constructs in NIH/3T3 cell. α-tubulin represented as cytosol marker and histone H3 as nuclear marker. (d) Immunofluorescence images showed that IFT 52 localization depending on Kat2b existence in the nucleus and function of acetylation. shControl NIH/3T3 cells transfected by empty vector, and stable Kat2b depleted NIH/3T3 cells, as shKat2b, transfected by empty vector and mutant Kat2b constructs. Nuclei were stained with DAPI (blue), transfected vector constructs were EGFP (green), acetylated α-tubulin and γ-tubulin were stained for primary cilia (red), and IFT52 were indicated yellow. White arrows indicate IFT components located in the cilia axoneme. The proportion of IFT52 distribution by categories are displayed below the graphs; ciliary axoneme, basal body, and not on the cilia. The graph on the bottom right indicated fluorescence intensity of ciliary IFT52. Scale bar: 2 μm.
