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. 2010 Nov 23;8(11):e1000545. doi: 10.1371/journal.pbio.1000545

Figure 2. eIF3f is involved in the deubiquitination of activated Notch.

Figure 2

HEK293T cells were transfected with: (A) ΔE, His-ubiquitin, two increasing doses of eIF3f shRNA #3 (indicated by + and +, respectively), and meIF3f as indicated; (B) ΔE, ΔE-LLFF, NIC or Notch full length (FL), His-ubiquitin, the larger dose of eIF3f shRNA used in (A) (+), and meIF3f as indicated; (C) ΔE, His-ubiquitin, shRNA #3, and two doses of meIF3f WT or Mut as indicated by a filled triangle. In all panels, cells were lysed under denaturing conditions and subjected to Nickel purification. Ni-purified material or WCE (5% of the total lysates) were resolved on SDS-PAGE and analyzed by Western blot using the antibodies indicated on the right of the panels. Notch p120 designates the membrane-anchored form of non-activated full-length Notch, resulting from furin cleavage. Overexpressed meIF3f has a higher apparent molecular weight than the endogenous human eIF3f because of its HA tags. White lines indicate that intervening lanes have been spliced out.