Skip to main content
. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Oncogene. 2010 May 24;29(30):4287–4296. doi: 10.1038/onc.2010.175

Figure 3. LMP2A Associates with p63α in Cytoplasm and Nuclear Membrane and Requires the PY and ITAM Signaling Motifs.

Figure 3

A Whole cell lysates from HEK293 cells transiently expressing pBabe+pcDNA3, ΔNp63α, LMP2A, or LMP2A+ΔNp63α were fractionated into cytoplasmic (cyto - C), nuclear (nuc – N), and nuclear membrane (NM) fractions. Protein expression in each fraction was determined by western blot. Data is also expressed graphically as % of the total amount of the protein of interest that was detected in each fraction. B Itch (Standard deviations were calculated based on n=2). C p63α (Standard deviations were calculated based on n=2). D LMP2A (n=1). E Localization of LMP2A-p63α-Itch complexes was determined by immunoprecipitation of p63α from fractionated lysates (cytoplasmic – C, nucleoplasmic – N, nuclear membrane – NM) followed by western blot for LMP2A and Itch. For beads-only controls, protein G beads were added to ΔNp63α and ΔNp63α+ LMP2A cytoplasmic fractions, without an antibody IP, to determine non-specific protein binding. F Requirement of LMP2A signaling motifs for association of LMP2 with p63α was determined by immunoprecipitation of LMP2A from HEK293 cells transiently expressing pBabe, LMP2A or the PY, ITAM, or YEEA mutants of LMP2A, followed by western blot for Itch and p63α. For beads-only controls, protein G beads were added to LMP2A lysates, without an antibody IP, to determine non-specific protein binding.