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Fig. 3a PRPS1 knockdown reduces NAD+ levels, NAD+/NADH ratio, and cell viability in HEI-OC1 cells. (a) Western blot analysis of PRPS1 expression after transfection with PRPS1-siRNA or negative-siRNA, with tubulin as the internal control. (b) Quantification of PRPS1 expression levels from (a). Measurement of intracellular (c) NAD+ levels and (d) NAD+/NADH ratio in si-CTRL and si-PRPS1 groups. Measurement of intracellular (e) NAD+ levels and (f) NAD+/NADH ratio in si-PRPS1 and si-PRPS1+NMN groups. (g) Cell viability assessed by CCK-8 assay. Data are presented as three independent experiments’ mean ± SD. ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001 compared with the si-CTRL; ####p < 0.0001 compared with the si-PRPS1.