FIG 3.

Tegument proteins pp71 and UL35 and immediate early protein IE1 downregulated ss NID1 levels. (A to C) HFFs were infected at the indicated MOI with Adpp65 (A), Adpp71 (B), or AdUL35 (C) or with HCMV at an MOI of 5 (V lane), harvested at 72 hpi, and assayed for the indicated tegument protein. Note that each blot used different lysate loading levels (A, 1 μL; B, 10 μL; C, 1 μL). The data in C also include cells infected with a UL35A-encoding retrovirus selected with puromycin, then reseeded and allowed to grow for 72 h. (D) HFFs infected with the indicated Ad vector at the indicated MOI were harvested at 72 hpi, and cell lysates (20 μL) were assayed for ss NID1. (E) UL35A retrovirus-infected HFFs were selected with puromycin, then reseeded and allowed to grow for 72 h. Cell lysates (20 μL) were assayed for ss NID1 levels. (F) HFFs were infected with Towne at an MOI of 5, AdIE1, AdIE2, or Adic (+Adtrans; all at an MOI of 10) and harvested at 72 hpi. Cell lysates (20 μL) were assayed via Western blotting for NID1, IE1 (monoclonal antibody p63-27), and IE2 (monoclonal antibody CH16.0). Pan-actin or lamin B was used as a loading control.