Figure 3
P-PantSAc rescues decreased acetylated lysine levels in dPANK/fbl RNAi treated cells and HoPan-induced larval lethality. (a) Control Drosophila S2 cells show normal levels of histone acetylation visualized by immunofluorescence using antibodies specific to acetylated lysines (AcLys in green). Rhodamin-phalloidin was used to visualize actin (F-actin in red), and DAPI was used to stain nuclei (in blue). (b) dPANK/fbl RNAi was used to downregulate levels of Drosophila PANK inducing a decrease of lysine acetylation levels. (c) Addition of P-PantSAc to the medium of dPANK/fbl RNAi-treated cells restored lysine acetylation levels. Bar represents 20 µm. (d) Western blot analysis for Acetylated Tubulin levels and Acetylated histone levels of control cells, dPANK/fbl RNAi treated cells and dPANK/fbl RNAi treated cells supplemented with P-PantSAc. An antibody against Acetylated lysines (AcLys) was used to detect Acetylated Tubulin and Acetylated histones. An antibody specific for Acetylated histone H3 was also used to detect levels of Acetylated Histone 3 (AcH3). Tubulin was used as a loading control. (e-g) Quantifications of (d). (For all graphs mean ± SEM is given. Two-tailed Student t-test was performed. *p < 0.05, **p < 0.01, ***p < 0.001). (h) HoPan provided via the food of developing Drosophila larvae induced lethality. Larval lethality was rescued by co-addition with HoPAN of CoA or P-PantSAc at the pupal stage. (For 3D mean ± SEM is given. Two-tailed Student t-test was performed. *p < 0.05, **p < 0.01, ***p < 0.001).
