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. Author manuscript; available in PMC: 2010 Feb 15.
Published in final edited form as: Dev Biol. 2008 Dec 10;326(2):347–356. doi: 10.1016/j.ydbio.2008.11.035

Figure 5. hid 5’F drives reporter GFP expression in imaginal discs.

Figure 5

(A) Schematic representation of hid enhancer constructs: Black lines indicate non-coding DNA, open boxes indicate reporter GFP and lacZ, “X” represents mutated E2F site. (B and C) Very low level of GFP reporter expression are observed in 3rd instar eye and wing discs from WT constructs (hid5’F-WT). (D and E) Mutation of the consensus E2F binding site dramatically upregulated GFP reporter expression in eye and wing discs. (F-K) Eye and wing imaginal discs 4-5 hr after gamma-irradiation (4Gy). (F and G) A high level of GFP reporter expression from hid5’F-WT was observed in most cells in the eye disc (F) and wing disc (G). (H and I) Acridine Orange staining of irradiated discs show increased cell death in the eye disc (H) as well as the wing disc (I). (J and K) Irradiation induced cell death in the eye disc (J) and wing disc (K) was suppressed by removal of hid. (L-N) Increased expression of hid reporter in rbf mutant background. In situ hybridizations of GFP reporter expression in WT and rbf15Δ/120 eye discs are shown in (L) and (M), respectively. Increased β-gal reporter level was also observed in rbf mutant clones (N, magenta, arrow). The position of the MF groove was indicated by white arrows. Anterior of eye disc is to the left and dorsal is up. Anterior of the wing disc is to the left and ventral is up. (O) Sequence of the conserved hid 5’ E2F site in Drosophila species. E2F binding site is underlined. E2F mutant sequence of mutated E2F binding site is shown at the bottom. Mutated bases are shown in lowercase letters. (P-S’) The hid 5’F responds various biological cell death stimuli. GFP reporter expression from whole third instar larva from hid5’F-WT (P) and hid5’F-E2FMut (Q). (R) A 42hr APF pupal retina showed high level of hid 5’F-WT GFP reporter expression in ommatidia at the periphery of the eye disc and in some interommatidial cells. (S-S’) High magnification image of 42hr APF pupal retina showed upregulation of GPF reporter expression from hid5’F-WT enhancer in some interommatidial cells.