Fig. 4: Targeting IL-6/STAT3 pathway impairs liposarcoma proliferation.
From: A metabolic crosstalk between liposarcoma and muscle sustains tumor growth
Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells supplemented with vehicle, anti IL-6 (BE8) A, or Siltuximab B, evaluating expression of serine synthesis pathway genes: Phgdh, Psat1 and Psph. (A n = 15, B n = 6, experimental replicates). C, Proliferation assay performed on IB115 cells alone or cocultured with C2C12 cells grown in media without Serine and Glycine and supplemented with vehicle or anti IL-6 (BE8). (n = 3 experimental replicates). D, Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells supplemented with vehicle or GP130 inhibitor (BZA), evaluating expression of serine synthesis pathway genes: Phgdh, Psat1 and Psph. (n = 15 experimental replicates). E qChIP experiments showing the relative amounts of mono-methylation of lysine 9 of histone H3 (H3K9Me1) on the Psat1 promoter in C2C12 treated or not with recombinant IL-6. Results were represented as the relative ratio between the mean value of immunoprecipitated chromatin (calculated as a percentage of the input) with the indicated antibodies and a control irrelevant antibody. (n = 3 experimental replicates). F NAD+/NADH ratio in C2C12 treated or not with recombinant IL-6. (n = 3 experimental replicates). G STAT3, and P-STAT3 protein expression assessed by immunoblot in C2C12 cells incubated 16 h with recombinant IL-6, IB115, or HPAC conditioned media. Tubulin was used as the loading control. H Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells (ratio 1:2) supplemented with vehicle or STAT3 inhibitor, evaluating expression of serine synthesis pathway genes: Phgdh, Psat1, and Psph. (n = 15 experimental replicates). I Proliferation assay performed on IB115 cells grown in media without Serine and Glycine and cocultured with C2C12 cells treated or not with STAT3 inhibitor. (n = 3 experimental replicates). Data are shown as means ± SEM and three or more independent experiments were performed. Statistical tests were all adjusted for multiple comparisons and included two-tailed unpaired t-test for (e, f), one-way (a, b, d, h) and two-way analysis of variance (ANOVA) for c and i. *P < 0.05, **P < 0.01, ***<0.001, n.s. non-significant. Source data are provided as a Source Data file.
