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. 2010 Jun 10;38(5):722–732. doi: 10.1016/j.molcel.2010.05.002

Figure 3.

Figure 3

ISG15 Conjugation to Proteins Not Previously Identified as ISG15 Targets

(A) Modification of human proteins not previously identified as ISG15 targets. HEK293T cells were transfected with plasmids expressing either TAP-E6AP or HA-p53 alone (lane 1), combined with Ube1L, UbcH8, Herc5, and ISG15 (lane 2), or combined with Ube1L, UbcH8, and Herc5 (lane 3). Cell extracts were prepared 48 hr posttransfection and analyzed by immunoblotting with anti-TAP or anti-HA antibodies. See also Figure S2.

(B) ISG15 modification to foreign proteins. HEK293T cells were transfected with plasmids expressing the indicated proteins and either V5-HPV18 L1 protein or TAP-tagged Shigella OSPG protein and analyzed as in (A) with anti-V5 or anti-TAP antibodies.

(C) Modification of nonoverlapping fragments of IQGAP1. A schematic of IQGAP1 is shown (top), along with three nonoverlapping fragments, A–C, with amino acid numbering shown. Plasmids expressing TAP-tagged fragments A–C were transfected into HEK293T cells and analyzed as in (A) using anti-TAP antibody.