Fig. 7: NEDDylation inhibition prevents SKP2 functions and suppresses tumor growth in vivo. | Nature Communications

Fig. 7: NEDDylation inhibition prevents SKP2 functions and suppresses tumor growth in vivo.

From: MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57Kip2 targeting

Fig. 7

a Representative western blot (n = 3 independent experiments) of the indicated proteins on RD and JR1 cells treated for 48 h with either vehicle (DMSO) or MLN4924, at the reported Growth Inhibition (GI) concentrations (refer to Fig. S7A for details). Vinculin is the loading control. b Growth curve analysis of RD and JR1 cells treated as in (a). n = 3 independent experiments, data presented as mean values ± SD, Student’s two-tailed t-test. c Representative western blot (n = 3 independent experiments) of the indicated proteins on RD and JR1 cells treated as in (a). Vinculin is the loading control. d Representative light microscopy pictures of β-Galactosidase staining (blue) of RD and JR1 cells treated with vehicle (DMSO) or GI50 MLN4924. Scale bar = 100 μm. e Histogram depicts the quantification of the percentage of senescent cells per field of RD and JR1 cells treated as in (d). n = 3 independent experiments, data presented as mean values ± SD, Student’s two-tailed t-test. f Histogram depicts the quantification of Caspase 3/7 activity at the reported MLN4924 concentration and time points calculated as fold increase over vehicle (DMSO) (n = 3 independent experiments, data presented as mean values ± SD, two-way ANOVA). g Images of RD tumors explanted from mice treated with either vehicle or MLN4924 for 3 weeks. h Tumor volume of RD xenografts treated as in (g) (vehicle n = 5, MLN4924 n = 5) assessed by caliper measurement represented in mm3 followed for 3 weeks treatment. Data presented as mean values ± SD, two-way ANOVA. i Tumor weight of transplanted RD xenografts treated as in (g). Box plots show 25th to 75th quartiles, black bar shows the median, and whiskers go down to the smallest value and up to the largest. One-way ANOVA. j Representative images (n = 3 independent experiments) of H&E, p27Kip1, p21Cip1, Cleaved Caspase and Ki67 immunohistochemistry of tumor sections from RD xenografts treated with either vehicle or MLN4924. Scale Bars = 100 μm. Source data are provided as a Source Data file.

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