Figure 2.

Missense p53 mutants in PDAC cells are stabilized by Hsp90. (A) Hsp90-dependent aberrant stabilization of mutp53 proteins in PDAC cell lines. Cells were treated for 24 h with the indicated concentrations of Ganetespib, Onalespib, or DMSO. One representative immunoblot out of three each is presented. HSC70, loading control. Total AKT (‘tAKT’, AKT serine/threonine kinase 1) as well-known Hsp90 client serves as functional control for an Hsp90 inhibition. (right) Diagrams represent the means ± SEM of densitometric quantifications of at least two independent experiments with technical replicates (total n ≥ 3 immunoblots), normalized to HSC70. Calculated relative to control DMSO treatments (con). (B) Cell confluence determination. Representative images of cells after treatment with 200 nM Ganetespib, Onalespib, or solvent control for 24 h. Cell confluency was analyzed using a Celigo imaging cytometer. Scale bars, 100 µm. Confluence was calculated relative to their respective DMSO control from n = 3 biological replicates. (A, B) Student’s t test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ns, not significant.