Fig. 3

LAP3 downregulates arginine level by interfering with ASS1 in BMECs. a-e Targeted metabolomics were performed to detect intracellular levels of citrulline (a), putrescine (b), agmatine (c), ornithine (d), and spermine (e) with IFN-γ or in combination with 100 mg/mL LAP3 inhibitor, bestatin. f Western blotting was used to detect the expression of ASS1 protein in LAP3 knocked-down BMECs upon IFN-γ treatment. Full-length blots are presented in Supplementary Fig. S7. g Western blotting showed the changes of ASS1 protein expression after treatment with LAP3 inhibitor bestatin in malignant transformed BMECs cells. Full-length blots are presented in Supplementary Fig. S8. h Determination of the change in arginine content of malignant BMECs treated with the LAP3 inhibitor, bestatin. The relative level of the target protein was estimated using densitometry and the ratio was calculated relative to the GAPDH control. Bars represent mean values with error bars to represent SD from three independent replicates. Differences between mean values were assessed by two-tailed Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001, compared to control