FIG 3.

Knockdown of Stx6 does not affect HCMV nuclear translocation in fibroblasts or epithelial cells. HEL fibroblasts (A and B) and ARPE-19 epithelial cells (B and D) were treated with siRNA targeting Stx6 or a scrambled (Scr) siRNA control. Treated cells were incubated for 24 h to ensure maximal knockdown. Immunoblots were performed on whole cell lysate and Stx6 band density was measured and normalized to quantify knockdown efficiency (B and D). The treated cells were then synchronously infected with HCMV TB40/E UL32-GFP (MOI = 5). Protein was harvested from infected and mock-infected fibroblasts (E and F) and ARPE-19 cells (G and H) at 3 hpi and 6 hpi, respectively. Lysates were immunoblotted for HCMV IE-1 and actin (E–H). Protein amount was quantified using LI-COR Image Studio software. Student’s t-tests were used to assess statistical significance. N = 3 per cell-type. (C) *, P < 0.05; **, P < 0.01; ***, P < 0.001.