Figure 5: PolII S2P binding, CRTC binding and histone acetylation are regulated by GCN5, Tip60 and CRTC.

(a,b) Histone acetylation and CRTC binding after spaced training are abundant in comparison with those after massed training at Bx and Smr gene loci. The purified MB nuclei expressing FLAG-KASH prepared from naive control flies or flies 1 day after spaced training or massed training were subjected to ChIP analysis using anti-H3K9Ac, anti-H4K16 or anti-CRTC antibodies. The ChIP DNA was analysed at Bx (a) or Smr (b). The naive control flies were used to calculate relative enrichment (n=3 for all data.). (c,d) PolII S2P binding, CRTC binding and histone acetylation are reduced by knockdown of GCN5, Tip60 and CRTC. Flies expressing FLAG-KASH in the MBs and carrying MBsw were crossed with the UAS-RNAi lines, as indicated at the bottom of each panel. The MB nuclei from naive control flies and flies 1 day after spaced training were collected as in a and subjected to PolII S2P ChIP assay. The ChIP DNA was analysed at Bx (c) or Smr (d). The naive control flies were used to calculate relative enrichment (n=3 for all data). *P<0.05 determined by Student’s t-test.