1. Proteolytically processed forms of Gli2 do not localize to cilia but the activator, repressor and zinc finger domains are dispensable for the ciliary localization of Gli2.
Proteolytically processed forms of Gli2 do not localize to cilia but the activator, repressor and zinc finger domains are dispensable for the ciliary localization of Gli2. (A) Gli2–GFP ESCs were stained for AcTub (red) and GFP (green). The processed forms of GFP-tagged Gli2 (Gli2-78 and Gli2-110) do not localize to cilia (red arrowheads), whereas GFP-tagged Gli2 lacking the N-terminal repressor domain, C-terminal activator domain or central zinc finger domain (Gli2ΔRep, Gli2ΔAct or Gli2ΔZF, respectively) robustly localize to the cilium (white arrowheads). Insets, the individual cilium indicated by the white arrowhead. Scale bar: 5 µm. (B) Gli2-78, Gli2ΔRep and Gli2ΔAct localize more robustly to the nucleus than wild-type Gli2–GFP. Scale bar: 5 µm. (C) Lysates from Gli2Rev/+ (Rev), Gli2Gli2–GFP/+ (WT), Gli2Gli2-110–GFP/+ (-110) and Gli2Gli2-78–GFP/+ (-78) ESCs were blotted for GFP and tubulin, or were immunoprecipitated with anti-GFP and blotted for GFP and Sufu. There is less Gli2-110 protein than wild-type or Gli2-78 protein. Neither Gli2-78 nor Gli2-110 co-immuoprecipitate Sufu. (D) Gli2ΔRep–GFP, Gli2ΔAct–GFP and Gli2ΔZF–GFP ciliary localization prevalence and pixel intensity were compared to Gli2–GFP. + denotes lowest pixel intensity, whereas ++++ denotes highest pixel intensity. *P<0.05 (Chi-squared test, as compared to WT).
Nicole Santos, and Jeremy F. Reiter J Cell Sci 2014;127:
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