Fig. 6: C1GALT1 is required for EWSR1::FLI1-mediated transformation.

a Soft agar assay monitoring colony formation of A673 cells stably expressing a NS, EWSR1, C1GALT1, or SMO shRNA. Data are presented as mean ± SD (n = 3 biologically independent experiments). P-values were calculated using one-way ANOVA with post-hoc Dunnett’s multiple comparisons test. b Tumor formation of A673 cells stably expressing a NS, C1GALT1, SMO, or EWSR1 shRNA following subcutaneous injection into NSG mice. Tumor dimensions were monitored every 3 days starting at day 3, when tumors became palpable. Data are presented as mean ± SD (n = 6 mice per group). P-values were calculated at day 24 using two-way ANOVA with post-hoc Tukey’s multiple comparisons test. c Representative IHC images showing Ki67, C1GALT1, SMO, GLI1, and FLI1 staining in tumors derived from mice in (b) at day 24. Scale bar, 50 µm. d Colony formation of NIH 3T3 cells expressing vector or EWSR1::FLI1 and an NS, C1GALT1, or SMO shRNA. Data are presented as mean ± SD (n = 3 biologically independent experiments). P-values were calculated using one-way ANOVA with post-hoc Tukey’s multiple comparisons test. e Colony formation of NIH 3T3 cells expressing vector, SMO or C1GALT1. Data are presented as mean ± SD (n = 3 biologically independent experiments). P-values were calculated using one-way ANOVA with post-hoc Dunnett’s multiple comparisons test. f Tumor formation of NIH 3T3 cells ectopically expressing SMO or C1GALT1, or cells ectopically expressing C1GALT1 and an SMO shRNA, following subcutaneous injection into NSG mice. Tumor dimensions were monitored every 5 days starting at day 5, when tumors became palpable. Data are presented as mean ± SD (n = 5 mice per group). P-values were calculated at day 30 using two-way ANOVA with post-hoc Tukey’s multiple comparisons tests. g Representative IHC images showing Ki67, C1GALT1, SMO, and GLI1 staining in tumors derived from mice in (f) at day 30. Scale bar, 50 µm. Source data are provided as a Source Data file.