Effects of vhs on the levels of partially complementary viral transcripts and dsRNA levels.

Abstract

A and B) Diagram of the UL23/UL24 (A) and UL30/UL31 (B) regions of the HSV-1 genome, with the transcripts detected by Tombacz et al. 2017 and earlier studies depicted. The probes used for northern blots analysis are diagrammed. Coordinates are from the HSV-1 strain 17 reference genome (X14112). C and D) HeLa cells were infected with HSV-1 (KOS) WT and Vhs- for 6 or 12 hours at an MOI of 10. Equal amounts of total RNA were analyzed by Northern blot with probes specific for either the UL23 (C) or UL24 (D) transcripts. Results shown are representative of 3 independent experiments. E) Detection of UL23/UL24 duplex RNA. HeLa cells were infected with HSV-1 (KOS) WT and Vhs- for 12 hours at an MOI of 10. Total RNA was extracted and digested with RNaseA/T1 (high salt buffer) or RNaseIII followed by RNaseA/T1. Undigested input (1/10) and digested samples were analyzed by Northern blot with a probe specific for the predicted overlap between the UL23 and UL24 transcripts. F) Detection of UL30/UL31 duplexRNA. The experiment was performed as in E) but with a probe specific for the predicted overlap between UL30 and UL31 transcripts. Results shown in E) and F) are representative of 4 independent experiments each.</p