Extended Data Fig. 7: Effects of LIF on HUVEC.
A. Representative histological images of subcutaneous tumours (scale bar 50 µm). B. Quantification of LIF positive areas in tumours using images from Fig. 7h. Results are expressed as the percentage of the LIF-positive area relative to the field area (n = 6, Scrambled 3 males and 3 females, LIF-KO C3 4 males and 2 females). C. Single-cell RNA-Seq average expression for LIFR in samples from previously untreated patients with NSCLC (n = 5) stratified by cell-type. Average expression is shown in each cell type without category aggregation. D. HUVEC cell proliferation measured by crystal violet assay after 24 h treatment with or without LIF (n = 3). E. Migration assay (6 h) quantification in HUVECs treated with increasing concentrations of glycosylated and non-glycosylated LIF, with increasing concentrations of FGF2 and VEGF as positive controls. Data is represented as fold change relative to control (media containing 0.5% of FBS only, n = 3). F. Migration assay (6 h) quantification in HUVECs treated with glycosylated and non-glycosylated LIF, with FGF2 and VEGF as positive controls. Data is represented as fold change relative to control (media only, n = 3 and n = 4 for 1% FBS). G-H. Representative images (G) and quantification (H) of tube formation assays in HUVECs treated with glycosylated and non-glycosylated LIF at 16 h. FGF2 and VEGF were included as positive controls. Unsupplemented HUVEC media was used as control. 20 ng/ml of glycosylated LIF were used instead of 10 to compensate for its double molecular weight. Quantification is expressed as fold change relative to control (media only, n = 3 for 0% FBS and n = 4 for 1% FBS). Dialyzed serum was used. I-J. Representative western blot (J) and quantification (I, n = 2) of pSTAT3 levels in HUVECs treated with glycosylated and non-glycosylated LIF at different concentrations (0, 1, 2.5, and 10 ng/mL). K. mRNA expression levels of genes related to VEGF in subcutaneous tumours from scramble and LIF-KO groups (n = 6). Graphs show average values and error bars represent the standard error of the mean (SEM). Statistical significance was determined by two-tailed Mann-Whitney unpaired test (B) or one-way ANOVA with multiple comparisons (I).
