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. 2010 May 26;299(2):F436–F444. doi: 10.1152/ajprenal.00674.2009

Fig. 5.

Fig. 5.

Nedd4–2 enhances ubiquitination of occludin. A and B: HEK293 cells transfected with FLAG-occludin (Occ), HA-ubiquitin, and Nedd4–2 and subjected to sequential immunoprecipitation and immunoblotting with an anti-HA antibody. Ubiquitinated occludin is seen as a broad smear and is bracketed. Representative experiment in A and pooled data quantified by densitometry shown in B. *P < 0.05 against Occ; #P < 0.05 against Occ + HA-Ub; Tukey's pairwise multiple comparison, means ± SE, n = 4. C and D: experiments similar to A and B but examining the effect of a catalytically inactive form of Nedd4–2 (Nedd4–2CS). Representative experiment in A and pooled data quantified by densitometry shown in B. Catalytic activity of Nedd4–2 is required for ubiquitination of occludin. *P < 0.05 by ANOVA against each of the other conditions.