Extended Data Fig. 1: Metabolic alterations by HSV-1 during lytic replication.

a, Heatmap of metabolites profiled in mock- and HSV-1-infected HepG2 cells at 12 hours post-infection (h.p.i) (MOI = 5). b, Cell viability of 293 T cells depleted with NAMPT, NMNAT1, NRK, or NADSYN, n = 3. c, Quantification of NAD⁺ and NMN in shNAMPT HeLa cells infected with HSV-1, supplemented with nicotinic acid (NA, 0.1 mM) or nicotinamide riboside (NR, 0.1 mM). d-f, Infection diagram (d) and HSV-1 titers (e, n = 3) in shCTL and shNAMPT HeLa cells with or without supplementation of NR, P = 0.0217 (no NR), P = 0.0028 (0.05 mM NR), P < 0.0001 (0.1 mM NR), P = 0.0005 (0.2 mM NR). Exogenous NR increases the abundance of the NAD⁺-related metabolites in sgNAMPT HeLa cells in a dose-dependent manner, n = 5 (f). Diagram was created with Biorender.com. g, HSV-1 titer in shCTL and shNAMPT HeLa cells without or with supplementation of nicotinamide riboside (NR, 0.1 mM) at 12 h.p.i. (MOI = 0.1), n = 3. h, Growth curve of HSV-1 is determined by intracellular virus in sgCTL and sgNAMPT HeLa cells, n = 4, P < 0.0001. Statistical significance was calculated using unpaired two-tailed t-tests, for d and f; two-way ANOVA for h. Data in b, d, f, h, and g are presented as mean values\(\pm\)SD. Graphics created with BioRender.com.