Fig. 2.

E4orf6-dependent ubiqutination and degradation of AAV large Rep is suppressed by the addition of a dominant-negative ubiquitin. (a) Immunoblots of large Rep immunoprecipitated with 7B73.2 and immunoblotted with anti-Flag (top panel) or anti-Rep (7B73.2) (bottom panel) in the presence or absence of the dominant-negative UBR7 or MG132 as indicated, and as described in the text. (b) Immunoblot of samples taken from 293 cells transfected with HIV LTR-driven large Rep alone (lane 1), or with co-expression of E4orf6 (lane 2) or E4orf6 together with UBR7 (lane 3). Large Rep was detected with 7B73.2 and E4orf6 expression was detected by HA (lower panel). Cellular tubulin was used as a loading control.