Fig. 2.

METTL14 redistribution increases as the HSV-1 replication cycle progresses. (A) NHDFs were infected with HSV-1 (MOI = 3) either in the presence (+PAA) or absence (−PAA) of 300 µg/mL PAA. Total DNA was collected at 3, 6, 9, 12, and 15 hpi and analyzed by qPCR using primers to the HSV-1 UL44 gene and human ribosomal protein 19 (RPL19) gene for normalization. (B) An equivalent infection time course was processed for an indirect immunofluorescence assay using antibodies to METTL14 or viral ICP8. Nuclear DNA was visualized using DAPI, and the boundary between the nucleus and cytoplasm is indicated by a dashed line. Note the accumulation of ICP8 into discrete vRCs at 6 and 9 hpi. (C) Biochemical fractionation and immunoblotting of HSV-1–infected cells collected over a time course of infection. Cell-equivalent cytoplasmic and nuclear fractions were probed using antibodies to METTL14, cytoplasmic β-tubulin, and nuclear histone H3.