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. 2011 Jul 15;25(14):1499–1509. doi: 10.1101/gad.2046211

Figure 1.

Figure 1.

(A) Western analysis of nuclear extract from Drosophila embryos probed with antibodies against Gcn5 and Ada2b. Extracts were isolated from OregonR (wild type, lane 1), UAS-Ada2bH1F2 in the absence of the GAL4 driver (lane 2), UAS-Ada2bH1F2 expressed under control of mef2-GAL4 (lane 3), and elav-GAL4 drivers (lane 4). (B) Lateral view of GFP expression in the ventral nerve chord and peripheral nervous system of a stage 15 elav-GAL4; UAS-Ada2bH1F2; UAS-GFP embryo. (C) Lateral view of GFP expression in somatic musculature of a stage 15 mef2-GAL4; UAS-Ada2bH1F2; UAS-GFP embryo. (D) GFP was plotted against yellow autofluorescence for single cells. GFP-positive nondebris events are shown within the green line (GFP+). The percentage of GFP-positive events is given in brackets beside each genotype. (E) Region map displaying the density of the SAGA ChIP-seq signal in muscle relative to neurons as IP/input (log2) for muscle-specific SAGA-bound genes (top panel), genes bound by SAGA in both muscle and neurons (middle panel), and genes bound by SAGA only in neurons (bottom panel). SAGA-binding density is shown for individual genes (rows) from −500 base pairs (bp) to +500 bp around the transcription start site (+1) in muscle (red) and neurons (blue) relative to the signal from a control Flag ChIP-seq experiment in untagged neuronal cells (green). (F) Binding profiles for SAGA (top panel) and acetylated H3-Lys9 (bottom panel) in muscle (red) and neurons (blue) are shown for the exba and wupA loci. The gene structure is indicated below the SAGA-binding profile: 5′ and 3′ UTRs are shown in green and white, respectively; constitutively and alternatively spliced exons are shown in black and red, respectively; and alternative transcription start sites are indicated by arrows. The Y-axis in the top panel represents the number of unique reads observed in two biological experiments for the SAGA ChIP-seq experiment for 20-bp windows across the two loci. The control profile in the top panel (green) represents the Flag ChIP-seq signal in untagged neuronal cells. ChIP for acetylated H3-Lys9 was performed on chromatin isolated from muscle and neurons. ChIP data represent mean percent input normalized to the coding region of a gene that is not expressed in embryos. Error bars denote standard error of the mean for three biological experiments.