Journal: Journal of Virology

Article Title: HSV-1 miRNAs are post-transcriptionally edited in latently infected human ganglia

doi: 10.1128/jvi.00730-23

Figure Lengend Snippet: Expression of ADAR1 and ADAR2 in human trigeminal ganglia. (A) Immunohistochemical analysis of ADAR1 and ADAR2 expressions in human TGs. Sections of paraffine-embedded TG tissue (4 µM) were stained with rabbit monoclonal α-ADAR1 IgG (Cell Signaling Technology), mouse monoclonal α-ADAR1 IgG (Santa Cruz Biotech), and mouse monoclonal α-ADAR2 IgG (Santa Cruz Biotech) and visualized using peroxidase-labeled polymer linked to goat α-rabbit and α-mouse immunoglobulins and 3,3′-diaminobenzidine. Slides were counterstained with hematoxylin and analyzed by an Olympus BX51 microscope equipped with a DP50 camera and CellF software (Olympus, Japan). The magnification is indicated in each panel (×100 or ×600). HE, ADAR1, and ADAR2 staining is shown in neurons; (B) RNA was extracted from human trigeminal ganglia (indicated TG samples TG1–TG5), and ADAR1 and ADAR2 transcripts were amplified using RT-PCR and specific primers. PCR products were visualized using by agarose gel electrophoresis. RTneg–control represents the samples processed without the reverse transcriptase. (C) Proteins were isolated from sections of two randomly selected human trigeminal ganglia (TG7 and TG9) with RIPA buffer [150-mM NaCl, 1% NP-40, 0.5% Na deoxycholate, 0.1% SDS, 50 mM Tris (pH 8.0), with protease inhibitors (cOmplete, Roche, Basel, Switzerland)] and analyzed by Western blot. ADAR1 was detected with the α-ADAR1 antibody (Santa Cruz Biotech) (upper panel) or the ADAR1 p150 antibody (Cell Signaling) (middle panel).

Article Snippet: ADAR1 was detected with the α-ADAR1 antibody (Santa Cruz Biotech) (upper panel) or the ADAR1 p150 antibody (Cell Signaling) (middle panel). . Low levels of HSV-1 miRNA editing during productive infection in culture We demonstrated that sequence-specific hyperediting of miR-H2 occurs during long-lasting latency in humans, which might have important implications for understanding the molecular mechanisms regulating latency.

Techniques: Expressing, Immunohistochemical staining, Staining, Labeling, Polymer, Microscopy, Software, Amplification, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Reverse Transcription, Isolation, Western Blot