Usp9x inhibition induces apoptosis in Kras mutant human cell line MIAPACA2, but Kras w/t BxPC3 cells remain unaffected.
(A) Immunoblot analysis shows that Usp9x KD in human mutant Kras PDAC cell line MIAPACA2 reduces mutant Kras expression levels, whereas Usp9x KD in the mouse mutant Kras cell line 8041 has no effect on Kras.
(B) Immunoblot analysis confirms Usp9x KD in human PDAC cell lines MIAPACA2 and BxPC3 with mutant Kras and w/t Kras, respectively. Usp9x KD in MIAPACA2 cells led to rapid apoptosis induction as indicated by PARP cleavage, but no such effect was observed in the w/t Kras cell line BxPC3.
(C) Phase contrast images further confirm apoptosis induction in MIAPACA2-Usp9x KD cells as compared to MIAPACA2-Control cells. No effect of Usp9x KD on the phenotype of BxPC3 cells grown in monolayer was observed.
(D) Representative column graph shows that BxPC3-Usp9x-KD cells show no effect on colony formation on 2% Matrigel in 3D culture when compared to BxPC3-Control cells. n.s., not significant.