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. 2008 Feb 18;205(2):465–477. doi: 10.1084/jem.20070725

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Copyright © 2008, The Rockefeller University Press

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Figure 4. — Substrates of CSN-regulated E3 ligases are dysregulated CSN5/JAB1^del/del thymocytes. Purified DN and DP/SP thymocytes from pools of 4 to 5 animals belonging to either WT or to each of the indicated CSN5/JAB1 knockout phenotypes (S, severe; Mo, moderate; Mi, mild; Fig. S5 shows phenotypic clustering of CSN5/JAB1^del/del thymocytes based on the DN:DP/SP ratio), likely resulting from incomplete penetrance of Lck-Cre-mediated recombination, were lysed and subjected to SDS-PAGE followed by immunoblotting with the indicated antibodies. (A) Immunoblot analysis for CSN5/JAB1, Cre, and Cul1 in purified DP/SP thymocytes. Cul1 is visualized as a doublet with the upper band representing the hyperneddylated isoform. (B) Immunoblot analysis for candidate substrates of CSN-regulated E3 ligases. Films were processed by densitometric scanning and normalized to values for β-actin for each lane to assess the relative values in each lane. Fig. S5 is available at http://www.jem.org/cgi/content/full/jem.20070725/DC1.