Fig. 8

In vivo experiment to validate the mechanism of LMOD1. A Diagram of the animal experiments. B Mouse xenograft tumors (n = 6 mice/group). C Xenograft tumor volumes. D Xenograft tumor’s weights at the end of the investigation. E mIF staining of FGF1 (green) and N-cadherin (green) proteins in mouse xenograft tumor stromal and parenchymal tissues. ACTA2 (pink), DAPI (Blue) and CK (red) for tissue-localization (Magnification, × 400, scale bars = 20 μm). F The diagram showing the procedures used in animal experiments in vivo. G Mouse xenograft tumors (n = 6 mice/group). H Xenograft tumor volumes. I Xenograft tumor’s weights at the end of the investigation. J mIF staining of N-cadherin (green) proteins in mouse xenograft tumor parenchymal tissues. DAPI (Blue) and CK (red) for tissue-localization. (Magnification, × 400, scale bars = 20 μm). K The association of LMOD1 mRNA expression with TIDE score as determined on the TCGA-CRC database (Spearman method, n = 620). L–N CRC tissue’s LMOD1 proteins IHC staining (Magnification, × 400, scale bars = 20 μm). O Bar plot showing the specific response rates for the high- and low-LMOD1 average H-score groups in 40 CRC patients. P Box plot illustrating the specific LMOD1 expression (H-Score) between non-responder and responder post anti-PD-1 therapy in 40 CRC patients (Wilcoxon test, *P < 0.05). Q Representative pictures of CT scan. Primary or metastatic tumor foci measured before initiation of immunotherapy (Baseline, BL). The red arrows indicate the primary or metastatic tumor foci. PD for progressive disease (PD), PR for partial response (PR). The expression level of LMOD1 can predict immunotherapy. R–T The co-stained LMOD1, CK, and CD8 images in the 3 immunophenotypes. Based on the spatial CD8 + T cells distribution, CRC tissues are categorized into 3 immunophenotypes, immune excluded, immune inflamed, and immune desert. Different sample measurements were taken. (Magnification, × 400, scale bars = 20 μm). Data are presented as the mean ± SEM ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. All experiments were repeated at least three times, independently