Extended Data Fig. 5: UGDH expression is a major determinant of LDL and HDL uptake in cancer cells.
From: Glycosaminoglycan-driven lipoprotein uptake protects tumours from ferroptosis
a. Proliferation (log2 doublings, 5 days) of indicated UGDH-deficient cell lines expressing a sgRNA resistant UGDH cDNA or an empty vector under treatment with the indicated concentrations of ferrous ammonium citrate (FAC). b. Immunoblot of TFRC (top) and IRP1 (bottom) in the indicated UGDH_KO cell lines expressing a sgRNA resistant UGDH cDNA or an empty vector treated or not with FAC (0.1 mg/mL). GAPDH included as a loading control. c. Immunoblot of GPX4 (left), SLC7A11 (centre), and AIFM2 (right) in the indicated UGDH_KO cell lines expressing a sgRNA resistant UGDH cDNA or an empty vector. GAPDH included as a loading control. d. Histograms of DiI-LDL uptake in Karpas299 UGDH_KO cells expressing sgRNA-resistant UGDH cDNA or vector control. e, f. Cellular uptake of DiI-labelled LDL (e) or HDL (f) in the indicated Karpas299 (left), A-498 (centre) and HeLa (right) UGDH_KO cells expressing a sgRNA resistant UGDH cDNA or an empty vector, quantified by median PE fluorescence intensity. g. Mass spectrometry analysis of isotope labelled ([3-13C]Cholesterol, left) and unlabelled (right) cholesterol in A-498 UGDH_KO cells expressing a sgRNA resistant UGDH cDNA or an empty vector cultured in the presence of native LDL (grey) or 13C-Cholesterol-labelled LDL for 4 h (dark blue) and 8 h (light blue). Data is presented as fold of the control LDL condition. a, e-g, Bars represent mean ± s.d.; a, e-g, n = 3 biological replicates. Statistics by two-sided unpaired t-tests compared to empty vector transduced cells (a, e-g). For gel source data, see Supplementary Fig. 1.
