Figure 5.

IFA demonstrating the CRL-mediated SAMHD1 loss during HCMV infection. (A) HF cells in chamber slides were mock infected or infected with HCMV (Towne) at an MOI of 1 for 72 h and double-label confocal IFA was performed with anti-SAMHD1 and anti-IE1 antibodies. FITC-labeled anti-rabbit IgG and rhodamine/Red X-coupled anti-mouse IgG were used for visualization. Hoechst stain was used to stain cell nuclei. Three single-labeled images and a merged imaged for SAMHD1 and IE1 are shown. Some virus-infected (arrows) and uninfected (arrowheads) cells were indicated. (B) HF cells were mock-infected or infected with HCMV for 96 h with DMSO or MLN4924 treatment for 24 h prior to cell fixation and double-label confocal IFA was performed as in (A). Inserts (a and b) are enlarged to indicate cells showing nuclear diffuse distribution of SAMHD1 (arrows) or aberrant nuclear distribution of SAMHD1 (arrowheads). Percentages (%) of cells showing different SAMHD1 distribution patterns in DMSO or MLN4924-treated cells are shown as graphs. The total cell numbers (n) counted are indicated.