Fig. 3

LIF represses CXCL9 through epigenetic silencing. a, b qRT-PCR analysis of the indicated genes in BMDMs. BMDMs were pre-incubated with 20 ng/ml LIF for 72 h and then stimulated with 5 ng/ml IFNγ or 10 μg/ml IL4 during 6 h (a) or with 0.1, 0.5, 1 and 5 ng/ml IFNγ for 24 h (b). c CXCL9 ELISA from BMDMs pre-incubated with 20 ng/ml LIF and then stimulated with 0.1 ng/ml IFNγ for 24 h. d CXCL9 ELISA from human CD11b⁺sorted cells (77% CD11b⁺ CD14⁺, see Supplementary Fig. 7b, c) from human GBM cultured with 20 ng/ml LIF for 72 h and then with 0.1 ng/ml IFNγ for 24 h. e ChIP of Tri-methyl-histone H3 (H3K27me3), EZH2 and acetyl-histone4 (H4ac) was performed in BMDMs treated with 20 ng/ml LIF for 72 h. Scheme shows the analysed CXCL9 promoter region. Representative data are presented as mean ± SD. f Representative images of IF of the indicated markers in human GBM organotypic slices (patients 1, 2, 3) incubated with 10 μg/ml anti-LIF for 3 days. Scale bar, 20 μm. (see Supplementary Fig. 8). Bottom, percentage of double positive cells relative to Iba1⁺ cells and percentage of CXCL9⁺ cells relative to the total number of cells. Data are mean of all patients ± SEM. Statistical analyses by Student’s t-test or Mann–Whitney T-test. *P < 0.05, **P < 0.01; ***P < 0.001; ****P < 0.0001