Fig 2: The nuclear interaction of α3-PML-IE1 is disrupted by digitoxin in HCMV-infected cells, and antiviral response genes are reduced. (A) Immunoprecipitation (IP) of PML was performed in HCMV-infected (MOI of 1), digitoxin (50 nM)-treated, or GCV (5 µM) cells at 24 hpi. IE1 and α3 were probed for analysis of their interaction with PML. The left panel shows the profile of different proteins in the nucleus (input), used for IP. (B) Reverse IP of α3 was performed in HCMV-infected digitoxin-treated cells at 24 hpi. IE1 and PML were probed to measure the status of α3-IE1-PML complex. The left panel shows the input used for the IP. (C–I) mRNA levels of IE1, MIEP-derived IE, IFN-β, CXCL-10, IL-1β, and IL-8 were measured in NI and I, digitoxin- and GCV-treated cells at 24 hpi. The experiments were performed in triplicates and repeated twice. ** represents p ≤ 0.01, and *** represents p ≤ 0.001. ns not significant.