Fig. 4: Scu increases IDH1 activity.
From: Scutellarin activates IDH1 to exert antitumor effects in hepatocellular carcinoma progression
A Proteome reactivity profile of HepG2 cells treated with Scu-B (100 μM). A protein affinity pull-down assay was subsequently performed. Then, the whole-cell lysates were pulled down with streptavidin beads, and the precipitated proteins were separated by SDS‒PAGE and processed by silver staining and LC‒MS/MS. B Labeling of recombinant IDH1 with Scu-B (100 μM). Immunofluorescence analysis of IDH1-Scu-B by Cy3-streptavidin. C, D CETSA confirmed the binding of Scu to IDH1. n = 3. E BLI assay showing the interaction of Scu with IDH1. F Schematic representation of the Scu activation of IDH1-mediated NADPH and α-KG production. G Scu increased α-KG production in HepG2 cells after treatment with Scu for 12 h. n = 3. H Quantitative analysis of the effect of Scu on the enzymatic activity of the IDH1 protein in HepG2 cells after treatment with Scu for 12 h. n = 3. I Scu dose-dependently activated the IDH1 protein in HepG2 cells. n = 3. J Scu had no influence on the level of D-2-hydroxyglutarate (D2HG) in HT1080 (IDH1-R132C) cells. n = 4. The data are mean ± SD; *P < 0.05; **P < 0.01; ***P < 0.001 compared to the control group.
