FIG 4.
SIM362–364 is required for the degradation of PML II but not for that of PML I. (A) No substantial changes in mycPML I or mycPML II mRNA level at −2 or 0 h in infections. HEp-2-TetOn cells expressing mycPML I or mycPML II were induced and infected with the viruses indicated. At −2 and 0 h, total RNA was extracted, cDNA was synthesized, and quantitative RT-PCR was performed to detect mycPML isoforms. The cycle threshold (ΔCt) number of mycPML was normalized against the 18S rRNA. (B and C) HEp-2-TetOn cells expressing mycPML I or mycPML II were infected with the viruses indicated in the presence or absence of CHX or MG132. Samples were taken for Western blotting (WB) with an anti-myc antibody (Ab) (B), and the half-lives of PML isoforms were calculated as described above (C). (D) The same samples shown in panel B parts a to f were probed with an anti-mCherry antibody in Western blot assays.