Fig. 1: TRIM23 regulates HSV-1 pathogenesis and autophagy in vivo.
From: TRIM23 mediates cGAS-induced autophagy in anti-HSV defense
a–e Trim23─/─ mice and wild type (WT) littermate controls of both sexes (8-week-old, sex-matched across treatments) were infected via intracerebral (I.C.) inoculation with 5 × 105 PFU of mutHSV-1. a Kaplan–Meier survival curves of mutHSV-1-infected Trim23─/─ and WT mice. b Viral titers in whole brain homogenates of Trim23─/─ and WT mice, determined by plaque assay at day 3 post-infection. c HSV-1 ICP-0, ICP-8, UL36 and gB transcripts in whole brain homogenates, determined by RT-qPCR at day 3 post-infection. d HSV-1 antigen expression in the indicated areas of mid-sagittal brain sections of mutHSV-1-infected Trim23─/─ and WT mice, determined at day 3 post-infection by immunohistochemistry. Scale bar, 3 mm (whole brain) or 50 µm (magnified areas). CP, caudate putamen; CB, cerebellum; HYP, hypothalamus; OB, olfactory bulb. e Endogenous p62 protein abundance in the whole brain lysates of mock-treated or mutHSV-1-infected Trim23─/─ and WT mice, determined at day 3 post-infection by immunoblot (IB). #1–#3 represent individual mice. f Viral ICP-0 and ICP-8 transcript expression in Neuro2a (mouse neuroblast) cells that were transfected for 30 h with the indicated siRNAs and then infected for 36 h with mutHSV-1 at the indicated MOIs, determined by RT-qPCR. g Endogenous p62 protein abundance and LC3B-I-to-II conversion in the whole cell lysates (WCLs) of Neuro2a cells that were transfected for 30 h with the indicated siRNAs and then either mock-treated or infected with mutHSV-1 (MOI 1, 5, and 10) for 24 h, determined by IB. h Viral titers in SH-SY5Y (human neuroblastoma) cells that were transfected for 30 h with the indicated siRNAs and then infected for 24 h with mutHSV-1 at the indicated MOIs, determined by plaque assay. i Endogenous p62 protein abundance and LC3B-I-to-II conversion in the WCLs of SH-SY5Y cells that were transfected for 30 h with the indicated siRNAs and then either mock-treated or infected with mutHSV-1 (MOI 0.2, 1 and 2) for 20 h, determined by IB. j Viral ICP-0 and ICP-8 transcript expression in primary human dermal fibroblasts (HDFs) that were transfected for 30 h with the indicated siRNAs and then either mock-treated or infected with mutHSV-1 (MOI 0.02) for 24 h, determined by RT-qPCR. k Endogenous p62 protein abundance and LC3B-I-to-II conversion in the WCLs of HDFs that were transfected for 30 h with the indicated siRNAs and then either mock-treated (–) or infected with mutHSV-1 (MOI 10) for 16 h, determined by IB. All data (a to k) are representative of at least two independent experiments (n = 7 mice for WT and n = 11 mice for Trim23─/─ (a); mean ± SD of n = 7 mice per condition (b); mean ± SD n = 8 mice for WT and n = 10 mice for Trim23─/─ (c); mean ± SD of n = 3 biological replicates (f, h and j)). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (Mantel–Cox log rank test (a), two-tailed Student’s t test with Welch’s correction (b, c), or two-tailed Student’s t test (f, h and j). Source data are provided as a Source Data file.
