ZFIN Figure: Finotto et al., 2023, Figure 8

Figure 8

LGALS1 is involved in suppression of the immune system

Volcano plot depicting differentially expressed genes in LBT003 and all other GSCCs (Left: downregulated genes in LBT003; right: upregulated genes in LBT003). y‐axis denotes −log₁₀(adjusted P‐value) while x‐axis shows log₂(avgFC) values. Cut‐offs were set at −log₁₀(adjusted P‐value) > 1.3 and abs(log₂(avgFC)) > 1.5.

Violin plot showing LGALS1 expression levels in GSCCs.

Representative double immunofluorescence images showing co‐expression of SOX2 (magenta) and GAL1 (cyan) in GBM tissue from LBT003 and LBT070. For enhanced visualization, a binary mask was generated from the SOX2⁺ cells and multiplied with the image of GAL1 staining in Fiji to exclude GAL1 staining in non‐tumor cells. Scale bars: 100 μm.

Mean fluorescence intensity values for GAL1 staining in SOX2⁺ cells in GBM tissue samples, normalized using Z‐scores within each sample. n = 6 tumor samples derived from five different patients.

Representative maximum intensity projections of a z stack of the head region of a Tg(mpeg1:mCherryF)^ump2; Tg(kdrl:lynEYFP)^md77 zebrafish embryo with a GFP‐labeled LBT070 LGALS1 KO tumor, at 1 dpi (left panel) and 5 dpi (right panel). Corresponding control images: see Fig 5I. GBM tumor cells are shown in green, GAMs in red, and blood vessels in blue. Scale bars: 50 μm.

Tumor volume at the start of 1 and 5 dpi time‐lapse movies (n = 12 and 9 (LBT070; 1 and 5 dpi), 12 and 5 (LBT070 LGALS1 KO; 1 and 5 dpi) zebrafish embryos; P = 0.0118 (LBT070), 0.0008 (LBT070 LGALS1 KO), 0.0927 (1 dpi), 0.9320 (5 dpi)).

Number of round GAMs at the start of 1 and 5 dpi time‐lapse movies (n = 12 and 9 (LBT070; 1 and 5 dpi), 12 and 5 (LBT070 LGALS1 KO; 1 and 5 dpi) zebrafish embryos; P = 0.1240 (LBT070), 0.0144 (LBT070 LGALS1 KO), 0.0053 (1 dpi), 0.1174 (5 dpi)).

Number of ramified GAMs at the start of 1 and 5 dpi time‐lapse movies (n = 12 and 9 (LBT070; 1 and 5 dpi), 12 and 5 (LBT070 LGALS1 KO; 1 and 5 dpi) zebrafish embryos; P = 0.1816 (LBT070), 0.0187 (LBT070 LGALS1 KO), 0.6851 (1 dpi), 0.0497 (5 dpi)).

Boxplot of GAM distance to the tumor of round GAMs within 30 μm of the tumor, at the start of 1 and 5 dpi time‐lapse movies (n = 137 and 48 (LBT070; 1 and 5 dpi), 291 and 39 (LBT070 LGALS1 KO; 1 and 5 dpi) GAMs; P = 0.0119 (LBT070), 0.9984 (LBT070 LGALS1 KO), 0.9999 (1 dpi), 0.0531 (5 dpi); boxes stand for 50% of the data and minima/maxima are indicated by the line ends).

Boxplot of GAM distance to the tumor of ramified GAMs within 30 μm of the tumor, at the start of 1 and 5 dpi time‐lapse movies (n = 29 and 13 (LBT070; 1 and 5 dpi), 40 and 20 (LBT070 LGALS1 KO; 1 and 5 dpi) GAMs; P = 0.8012 (LBT070), 0.9647 (LBT070 LGALS1 KO), 1.0000 (1 dpi), 0.9675 (5 dpi); boxes stand for 50% of the data and minima/maxima are indicated by the line ends).

Data information: The zebrafish experiments describe biological replicates, i.e. individual zebrafish embryos. The xenograft experiment was replicated twice for LBT070 LGALS1 KO, and three times for LBT070. In (F–H), data are presented as mean ± SD, and the P‐values were calculated by mixed‐effects model, followed by Šídák's multiple comparisons correction. In (I and J), the P‐values were calculated by two‐way ANOVA, followed by pairwise testing with Tukey's multiple comparisons correction. ns ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001.

Source data are available online for this figure.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO Mol. Med.