Fig 2: Baseline IL-11 concentrations in plasma controls in preclinical species and healthy human.

Fig 3: CRTC2 and SIK are key signaling nodes regulating LKB1-dependent functions. (A) Schematic cartoon of LKB1 family substrates. (B and C) IL-11 levels were quantified by ELISA from the supernatants of MC38 cells with indicated genotypes. EV = empty vector. For normalization, the LKB1-KO value of each experiment was set to 100%. Data are presented as the mean ± SEM. Statistical difference was assessed with a one-way ANOVA with Dunnett’s multiple group comparison post hoc test. (D) Tumor volumes (TVol) of MC38 cells with indicated genotypes were treated with either isotype control or anti-PD-1 antibody. Vertical dotted lines define the treatment time-points. EV = empty vector. Data are presented as individual tumor growth. TGDIso-500 is the tumor growth delay to reach 500 mm3 for each individual tumor in the isotype treatment group. (E) Quantification of intratumoral immune populations from isotype-treated tumors with indicated genotypes (depicted as percentage of CD45+ cells). Data are presented as the mean ± SEM; statistical difference was assessed by using one-way ANOVA with Sidak’s multiple group comparison post hoc test. (F) Scatter plots representing qRT-PCR of selected genes on cancer cells isolated from isotype-treated tumors presented in (D) and LKB1-WT tumors in Fig. 1D. Individual points represent different tumors, and the bar represents the median value. For normalization, the mean of the LKB1-KO group was set to 1. Statistical difference was calculated by one-way ANOVA with Dunnett’s multiple group comparison test. (G) Violin plot representing the time for MC38 tumors with indicated genotypes to reach 500 mm3. EV = empty vector. Statistical difference was assessed with a one-way ANOVA with Sidak’s multiple group comparison test. (H) Tumor volumes from A549 (Top) and H2122 (Bottom) cells transduced with inducible shRNA against CRTC2 and measured in mice treated with 10% sucrose or doxycycline. Data are presented as the mean ± SEM. Statistical differences in tumor volumes (TVol) at the end of treatment were assessed by using an unpaired t test.

Fig 4: LKB1 loss induces extensive transcriptional reprogramming. (A and B) Volcano plots showing the comparisons of LKB1-KO and LKB1-WT cell lines (A), and cancer cells isolated from LKB1-KO and LKB1-WT MC38 tumors (B). x-axis represents the logarithmic 2 scaled fold-change (FC) between the average of indicated samples; y-axis depicts the negative logarithmic 10 scaled P-value. Genes are colored based on significant Log2(FC) (P-value < 0.05). (C) IL-11 levels were quantified by ELISA from the supernatants of indicated cell lines. For normalization, the mean of the LKB1-KO group of each cell line was set to 100%. Data are presented as the mean ± SEM. Statistical difference was assessed by the t test. (D) Scatter plot representing IL-11 mRNA levels (left y-axis and black triangles) and tumor volumes at the end of treatment (EoT) (right y-axis and colored circles/squares) for tumors in Fig. 1D. X-axis groups data according to the genotypes. (E) Dot plot representing IL-11 mRNA levels in STK11 wild-type (WT) and mutant (MUT) NSCLC cell lines from CCLE (24). Statistical difference was calculated by using the Mann–Whitney U test. (F) Dot plot representing IL-11 mRNA levels from in silico deconvoluted cancer cells in STK11 wild-type (WT) and mutant (MUT) tumors using TCGA LUAD dataset. Statistical significance was tested by Wilcoxon-test. (G and H) mRNA quantified by qRT-PCR (G) and protein (quantified by ELISA) (H) for IL-11 levels from the STK11 mutant (MUT) and wild-type (WT) isogenic pair of A549 cells. Data are presented as the mean ± SEM. Statistical difference was calculated by the t test.