Supplemental Figures

Supplemental Figure 1 A B 140 120 WT IRF3 KO 100 120 80 60 100 40 cGAS 20 % Relative infectivity 80 0 IRF1 ZAPS cG AS MICB FUT4 TLR3 MDA5 60 MYD88MCOLNDDX60 MAP3K14 C 200 40 TRIM25 WT 150 IRF3 KO % Infectivity IRF3 KO cells 20 100 IRF7 50 0 infectivity % Relative 0 50 100 150 200 250 0 IDO % Infectivity WT cells IRF7 AIM2 RIPK2 MDA5 TRIM25 TNF RSF 10A Supplemental Figure 1. (A) Direct comparison of relative HCMV primary replication for each individual ISG between wild type and IRF3 KO fibroblast cells. Blue box = ISGs that reduced virus production by more than 2-fold in wild type cells, but were not in IRF3 KO cells. The green box = ISGs that reduced virus production by more than 2-fold in both wild type and IRF3 KO cells. Relative primary replication and virus production levels of HCMV for the ISGs in the blue box (B) and for ISGs that reduced primary replication by more than 50% (C) were plotted. Supplemental Figure 2 A B Supplemental Figure 2. Efficient knockdown of ZAP and TRIM25 by siRNA. Fibroblast cells were transfected with siRNA targeting ZAP, TRIM25 or a negative control siRNA. Total protein was harvested two days post transfection and levels of ZAP and TRIM25 determined by Western blot analysis. Supplemental Figure 3 A B Supplemental Figure 3. CpG and GpC corrected ratios for ORFs of alpha-herpesviruses. Corrected CpG and GpC ratios were calculated for alpha-herpesviruses as described in the materials and methods and in figure 3. Supplemental Figure 4 A B Supplemental Figure 4. CpG and GpC corrected ratios for ORFs of gamma-herpesviruses. Corrected CpG and GpC ratios were calculated for gamma-herpesviruses as described in the materials and methods and in figure 3. Supplemental Figure 5 A Corr_CpG: Betaherpesvirinae B Corr_GpC: Betaherpesvirinae 2.0 Cytomegalovirus Muromegalovirus Roseolovirus Unassigned 2.0 Cytomegalovirus Muromegaloviru Roseoloviru Unassigned Proboscivirus s Proboscivirus s 1.6 1.6 1.5 1.5 1.0 1.0 IE2 IE2 IE2 IE1 IE2 IE3 IE1 IE2 IE2 IE1 0.6 IE2 0.6 IE3 IE2 0.5 IE1 IE2 IE1 0.5 IE1 IE1 IE1 IE3 IE1 IE1 IE2 IE IE2 1 IE1 IE2 IE1 IE1 0.0 IE1 IE1 0.0 Suid betaherpesvirus 2 Suid betaherpesvirus 2 Murid betaherpesvirusMurid betaherpesvirusMurid 1 betaherpesvirus 2 8 Murid betaherpesvirusMurid betaherpesvirusMurid 1 betaherpesvirus 2 8 Aotine betaherpesvirusHuman 1 betaherpesvirusPanine 5 betaherpesvirus 2 Human betaherpesvirus 7Caviid betaherpesvirusTupaiid betaherpesvirus 2 1 Aotine betaherpesvirusHuman 1 betaherpesvirusPanine 5 betaherpesvirus 2 Human betaherpesvirus 7Caviid betaherpesvirusTupaiid betaherpesvirus 2 1 Macacine betaherpesvirus 3 Human betaherpesvirusHuman betaherpesvirus 6A 6B Macacine betaherpesvirus 3 Human betaherpesvirusHuman betaherpesvirus 6A 6B Saimiriine betaherpesvirus 4 Elephantid betaherpesvirus 1 Saimiriine betaherpesvirus 4 Elephantid betaherpesvirus 1 Cercopithecine betaherpesvirus 5 Cercopithecine betaherpesvirus 5 Supplemental Figure 5. CpG and GpC corrected ratios for ORFs of beta-herpesviruses. Corrected CpG and GpC ratios were calculated for beta-herpesviruses as described in the materials and methods and in figure 3. Supplemental Figure 6 A B C ISG20 MDA5 RIG-I 250 70 300 60 200 250 50 200 150 40 150 100 30 100 20 50 10 50 0 0 0 Relative expression (delta CT) Relative expression (delta CT) Relative expression (delta CT) UN 24HPI UN 24HPI UN 24HPI si NE G si TRI M25 siNEG siTRIM25 siNEG siTRIM25 Supplemental Figure 6. TRIM25 knockdown does not inhibit ISG induction. Wild-type fibroblast cells were transfected with TRIM25 siRNA or negative control siRNA. 48 hours later, the cells were then infected with TB40E-GFP at an MOI of 3. Total RNA were harvested at 24 hours post infection and the RNA levels of (A) ISG20, (B) MDA5 and (C) RIG-I were determined by Quantitative RT-PCR analysis. The result demonstrates that ISG induction following HCMV infection were not inhibited by TRIM25 knockdown..

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