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. 2016 Mar 6;7(14):18851–18864. doi: 10.18632/oncotarget.7944

Figure 6. Trastuzumab-resistant SKBR3 cells are sensitive to BAG-1 inhibition.

Figure 6

(A) SKBR3 parental or trastuzumab-resistant cells were treated over 5 days with DMSO (0.5% v/v), trastuzumab (32 μg/ml), Thio-S (50 μM), Thio-2 (50 μM), or combinations of these compounds as indicated. Cell viability was measured using crystal violet assay and is expressed as a percentage of DMSO control. Data shown on the graph are the mean ± SEM from 3 independent experiments, each with three technical replicates. Statistical significance was determined using two-way ANOVA with Holm-Sidak multiple comparisons test. *p < 0.05. (B) A representative immunoblot shows BAG-1 and HER2 expression levels in SKBR3 parental and trastuzumab resistant cells 4 days following siRNA (50 nM) knockdown. Densitometric analysis of immunoblots from 3 independent experiments was used to determine expression of total BAG-1 protein and is expressed as a percentage of untreated SKBR3 parental cells. The amount of BAG-1 protein knockdown by siBAG-1 is expressed as a percentage of the corresponding NSC treatment for each cell line; β-actin was used as a control for loading. (C) Bar graph shows the effect of BAG-1 knockdown by siRNA (50 nM) on the long-term growth potential of SKBR3 parental or trastuzumab-resistant cells. Data shown on graphs are the mean ± SEM from 3 independent experiments, each with 3 technical replicates. Unpaired t-test was used for comparison between different treatments; **p < 0.01.