Extended Data Fig. 10: Cell-surface sulfated GAGs promote lipoprotein uptake.
From: Glycosaminoglycan-driven lipoprotein uptake protects tumours from ferroptosis
a, b. Total cell surface HS (a) or CS (b) measured in A-498 cells treated or not with heparinases (0.1 U/mL) or chondroitinase (0.1 U/mL). c, d. Fold change in DiI-labelled LDL (c) and HDL (d) uptake of A-498 cells upon treatment with heparinases (0.1 U/mL), chondroitinase (0.1 U/mL), or both (blue), relative to uptake of untreated cells. e. Total cell surface hyaluronic acid (HA) measured in A-498 UGDH_KO cells expressing an UGDH cDNA or empty vector. f. Total cell surface HA (left), HS (middle), and CS (right) measured in A-498 cells treated or not with hyaluronidase (5 U/mL). g. Fold change in DiI-LDL uptake of A-498 cells treated with hyaluronidase (5 U/mL) or combined heparinases/chondroitinase (0.1 U/mL) relative to uptake of untreated cells. h. Representative image of HS immunostaining in A-498 cells treated or not with combined heparinases/chondroitinase (0.1 U/mL). Nuclei stained with DAPI. Scale bars, 20 μm. i. Representative image of Laurdan staining in A-498 cells treated or not with combined heparinases/chondroitinase (0.1 U/mL). Generalized polarization (GP) values represent membrane rigidity (higher value, red) or fluidity (lower value, blue). j. Histogram of GP values from images from experiment in (i). k. Fold change in pHrodo-transferrin uptake in A-498 cells treated or not with combined heparinases/chondroitinase (0.1 U/mL). l. Histogram of DiI-LDL uptake in cell suspension of 786-O tumours resected from mice treated or not with combined heparinases/chondroitinase. m. Uptake of DiI-LDL in the tumour suspension in (l). n, o. Representative IHC images (n) (20x magnification) and H-score quantification (o) of CD45 in Karpass299 tumours injected daily with HBSS or combined enzymes (10 U/mL). Scale bars, 100 μm. a-g, k, m, o: Bars represent mean ± s.d.; a-g, k, m: n = 3 biological replicates; o: n = 25 representative fields. Statistical significance determined by two-sided unpaired t-tests as indicated or compared to untreated cells (a-d, f-g, k, m, o) or empty vector expressing cells (e).
