Skip to main content
. 2014 Mar 28;24(5):542–559. doi: 10.1038/cr.2014.37

Figure 5.

Figure 5

BEZ235 stabilizes ErbB2 protein and lapatinib + BEZ235 combination overcomes resistance to Ab + lapatinib/Ab + BEZ235 in mouse primary cells. (A) Western blot analysis of cell lysates from BT474.Par and BT474.LapR cells grown in 3-D culture under different treatments for a week. (B) Immunofluorescence staining of BT474.LapR cells grown in 3-D culture under different treatment conditions for a week. Scale bar indicates 50 μm. (C) Protein stability assay in BT474.LapR cells treated with DMSO or BEZ235 (for a week) in combination with CHX over 24 h. (D) Quantification of ErbB2 signal intensity from C. (E) WST-1 proliferation assay after 72 h treatment with the indicated drugs or combinations in primary cells (254F.A + L/A + B.R and 315F.A + L/A + B.R) derived from PTEN−/−/NIC mice treated with Ab + lapatinib for 4 weeks and then with Ab + BEZ235 for 4 weeks, by which time resistance had developed. ***P < 0.001, by one-way ANOVA. (F) Western blot analysis of 254F.A + L/A + B.R and 315F.A + L/A + B.R primary cells after the indicated treatment for 3 h.