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. 2018 Oct 9;8:15005. doi: 10.1038/s41598-018-33379-x

Figure 2.

Figure 2

ICP27 interacts with UIF in co-immunoprecipitation assays. (A) HeLa cells were transfected with Flag-tagged UIFFL, UIFNT, UIFCT or Flag-tagged pUC18 plasmid DNA as indicated. Cells were infected 24 h after transfection with WT HSV-1 or were mock infected and 8 h later cell lysates were immunoprecipitated with anti-ICP27 antibody. Western blots were probed with anti-Flag antibody. Samples of each lysate were analyzed in parallel with the immunoprecipitated samples, and the Western blot is labeled input. The blot was probed with anti-YY1 antibody as a loading control. (B) Cells were transfected with Flag-tagged UIFFL plasmid DNA and were either mock infected or infected with WT HSV-1 KOS or WRL, in which the ALYREF binding site is mutated, as indicated, and immunoprecipitation was performed with anti-ICP27 antibody. Western blots were probed with anti-Flag, anti-ICP27 and anti-YY1 antibody as indicated. (C) Cells transfected with UIFFL plasmid DNA were mock infected or infected with HSV-1 KOS or ΔRGG, in which the RGG box RNA binding domain is deleted. Cells lysates were either treated or were not treated with RNase as indicated. Immunoprecipitation was performed with anti-ICP27 antibody and western blots were probed with anti-Flag antibody.