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. 2025 May 14;16:4489. doi: 10.1038/s41467-025-59774-3

Fig. 2. Functional characterization of CtCGT and CtF6H.

Fig. 2

a Catalytic functions of CtCGT and CtF6H in vitro. HPLC chromatograms of enzyme catalytic products of CtCGT using 1 (<i>) or 2 (<ii>) as sugar acceptor and UDP-Glc as sugar donor, and of CtF6H using 3 (<iii>) as acceptor and NADPH as donor. The ultraviolet absorption wavelengths for 1/2 and 3 were 300 and 340 nm, respectively. 1a (nothofagin), 1b (3′,5′-di-C-glycosylphloretin), 2a (2-hydroxynaringenin mono-C-glycoside), 2a-1 (isovitexin), 2a-2 (vitexin), 3a (scutellarein). b The workflow for VIGS experiments in C. tinctorius. c LC/MS analysis of VIGS samples, showing parallel reaction monitoring (PRM) chromatograms of HSYA, and (-)-ESI-MS/MS spectrum of [M-H]- ion at m/z 611. d The contents of HSYA in C. tinctorius upon VIGS treatment (n = 10, ten biologically independent samples were tested; The data were presented as mean values ± SD; Statistical significance was analyzed using a two-tailed t-test.). e Expression levels of CtF6H, CtCGT, and Ct2OGD1 in the EV and VIGS groups (n = 10, ten biologically independent samples were tested; The data were presented as mean values ± SD; Statistical significance was analyzed using a two-tailed t-test.). The source data underlying Fig. 2d, e are provided in a Source Data file.