Table 1

Analysis of PML colocalization with ICP0 during wt HSV-1 infection in the presence of MG132a

CellMean absolute no. ± SD% Foci
PMLICP0Co-locICP0+PML dots/ICP0 total dotsICP0+PML dots/PML total dots
HA+shLuci (control)39.6 ± 9.539.2 ± 9.736.9 ± 9.594.1393.18
HA+shPML (HALP)0.4 ± 0.941.8 ± 120.4 ± 0.90.95100
HALP+PML.I29.1 ± 10.729.8 ± 11.527.9 ± 10.693.6295.54
HALP+PML.II42 ± 12.445.1 ± 11.838.1 ± 12.184.4791.14
HALP+PML.III8.2 ± 3.640 ± 10.26.8 ± 31782.92
HALP+PML.IV23.9 ± 449.7 ± 13.723.3 ± 4.246.8893.57
HALP+PML.V9.5 ± 4.734.5 ± 69.5 ± 527.53100
HALP+PML.VI8.1 ± 2.636.3 ± 9.67.2 ± 2.419.8387.8
HALP+PML.I.KK25.5 ± 11.227.6 ± 11.525.4 ± 11.292.0399.5
HALP+PML.I.4K26.8 ± 7.8728.7 ± 9.225.5 ± 8.188.8595.15
HALP+PML.I.4K.Δexon97.7 ± 5.235.7 ± 14.52.4 ± 1.96.7231.48
HALP+PML.I.BB131 ± 8.431.6 ± 930.1 ± 8.595.2597
HALP+PML.I.BB222.8 ± 9.323.9 ± 9.822.2 ± 8.892.8898.6
HALP+PML.I.DCC12.5 ± 1135 ± 1212.5 ± 1135.71100
HALP+PML.I.ΔRING30.11 ± 8.732.6 ± 930.11 ± 8.792.3699.26
HALP+PML.I.MSIM21.8 ± 5.823.5 ± 6.121 ± 5.889.3696.33
HALP+PML.II.KK26.1 ± 939.5 ± 9.13.6 ± 3.99.1113.87
HALP+PML.IV.KK5.6 ± 322.7 ± 92.2 ± 29.6939.28
HALP+EYFP0.2 ± 0.539 ± 90.2 ± 0.55.13100
HALP+EYFP-PML.I.C-term19.9 ± 6.123.7 ± 6.219.4 ± 6.181.8597.48
HA+shPML.I38.08 ± 10.659.33 ± 12.637.83 ± 10.663.7699.34
  • a Cells expressing control (shLuci), anti-PML.I (shPML.I), or anti-PML shRNAs (shPML) and shPML cells expressing EYFP-PML isoform I, II, III, IV, V, or VI fusion proteins, mutant forms of EYFP-PML.I, -PML.II, or -PML.IV, EYFP-PML.I.C-term, or EYFP on its own were treated with MG132 and infected at an MOI of 2. After 4 h, the control and shPML and shPML.I cells were costained for ICP0 and endogenous PML. EYFP-PML was detected in shPML cells expressing the various EYFP-PML fusion proteins by autofluorescence. Single plane projections of short z-stacks of at least 10 cells were used for quantification of the mean absolute number of PML, ICP0, and colocalizing (Co-loc) foci per nucleus. The percentage columns indicate (i) the proportions of ICP0 foci that are associated with PML compared to the total number of ICP0 nuclear dots observed and (ii) the percentage of PML foci that are associated with ICP0 dots compared to the total of nuclear PML dots observed.