Fig. 1: Neuron-specific miR-9 represses HSV-1 replication and gene expression in Neuro-2a cells.

a Diagram of luc-HSV-1 virus and the procedure of screening for neuron-specific miRNAs regulating HSV-1 infection. b Neuro-2a cells in 96-well plates were transfected with 20 nM miRNA mimic for 24 h, then infected with luc-HSV-1 (MOI = 0.3) for 72 h before luciferase assays. The adjusted P values: miR-9-5p, <0.0001; miR-340-5p, 0.0002; miR-495-3p, <0.0001; miR-138-5p, 0.0003; miR-153-3p, 0.0007; miR-411-5p, 0.0021; miR-455-5p, 0.0035; miR-323a-3p, 0.0029; miR-338-3p, 0.0030; miR-330-3p, 0.019. c RT-qPCR analysis of miR-9 in the indicated tissues (left) or cells (right). d Neuro-2a cells were transfected with 20 nM miRNA mimic for 24 h, then infected with KOS (MOI = 0.3) for 48 h before viral titration. e The indicated cells were treated or not treated with 1 μg/ml Dox for 48 h, then infected with KOS (MOI = 0.3) for 48 before virus titration. f The indicated cells maintained in 1 μg/ml Dox were transfected with 80 nM antagomir for 24 h, then infected with KOS (MOI = 0.3) for 48 h before viral titration. g Neuro-2a cells were transfected with 20 nM miRNA mimic, then infected with KOS (MOI = 1) for 4 h before RT-qPCR analysis of the indicated viral transcripts. h N2A-C or N2AmiR9 cells maintained in 1 μg/ml Dox were infected with HSV-1 (MOI = 1) for 4 h before ChIP-qPCR analysis of the enrichment of H3K9me3 (left) or H3K27me3 (right) at the indicated promoters. NC, nonspecific control. n = 3 (c, e, g, h) or 4 (b, d, f) biologically independent samples. Data were analyzed by one-way ANOVA with Dunnett’s multiple comparisons tests (b, d), two-way ANOVA with Sidak’s multiple comparisons tests (e, f, h) or two-sided unpaired t tests (g) and are presented as the mean ± s.d. Source data are provided as a Source Data file.