Fig. 6: ROBO1 exhibits the characteristics of a dependence receptor.
a–c Representative CT combined with 3D organ reconstruction bioluminescence imaging displaying the outgrowth ability of Kpc1199 cells (b) or Panc02 cells (c) in mouse models: Premetastatic niche (PMN) (upper) and macrometastatic niche (MMN) (lower) (n = 5 mice per group, mean ± SEM.; two-tailed unpaired t test). Scale colour bars: 2.00 × 105−2.00 × 107. d, e Survival analysis of intrasplenic mouse models bearing Kpc1199 or Panc02 injection (e) (n = 8 mice per group). f Viability of SW-1990 cells with (+rSLIT2) or without (+PBS) 30 nM rSLIT2 exposure (n = 2 biological replicates, mean ± SEM., one-way repeated-measures ANOVA). The Y-axis represents OD values at 450 nm. (g) Viability of SW-1990ROBO1-FL cells exposed to 30 nM rSLIT2 with or without ROBO1 neutralizing antibody (n = 2 biological replicates, mean ± SEM., one-way repeated-measures ANOVA). The Y-axis represents OD values at 450 nm. (h) Colony formation assays evaluating the outgrowth ability of SW-1990 cells expressing ROBO1-FL or ΔROBO1 exposed to 10 nM or 30 nM rSLIT2 (n = 2 biological replicates, mean ± SEM.; two-tailed unpaired t test); (i) Tumour sizes in subcutaneous xenograft models utilizing PANC-1shCTRL or PANC-1 shROBO1 cells (n = 2 biological replicates, n = 7 mice per group, tumour volume was calculated as volume = 0.5 × length × width2, mean ± SEM., one-way Repeat-Measure ANOVA). j Tumour growth in subcutaneous xenograft models utilizing CAPAN-1CTRL or CAPAN-1SLIT2-oe cells (n = 2 biological replicates, n = 6 mice per group, tumour volume was calculated as volume = 0.5 × length × width2, mean ± SEM., one-way repeated-measures ANOVA). k Apoptosis of SW-1990 measured by flow cytometry with dual PI and Annexin V staining (n = 2 biological replicates, n = 3 tests per group, mean ± SEM.; two-tailed unpaired t test). (l) TUNEL assays performed on SW-1990CTRL, SW-1990ΔROBO1 and SW-1990ROBO1-FL cells with or without 30 nM rSLIT2 exposure (n = 3 biological replicates, 3 fields assessed per sample, mean ± SEM., two-tailed unpaired t test). TUNEL staining, green; DAPI, blue. Scale bars: 50 μm. (m) Caspase-3/7 activity in SW-1990 cells measured 48 h after serum starvation (n = 3 biological replicates, n = 5 tests per group, mean ± SEM., two-tailed unpaired t test). n Tumour growth in subcutaneous xenograft models utilizing SW-1990CTRL, SW-1990ΔROBO1 and SW-1990ROBO1-FL cells measured every 4 days (n = 6 mice per group, tumour volume was calculated as volume = 0.5 × length × width2, mean ± SEM., one-way Repeat-Measure ANOVA). Source data are provided in the Source Data file.
