Supplemental Figures and Legends from Mechanisms of Acquired Resistance to Trastuzumab Emtansine in Breast Cancer Cells

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Supplemental Figure 1. Upregulated BHLHE41 in KPL-4 TR and BT-474M1 TR cells does not confer resistance to T-DM1. Supplemental Figure 2. Flow cytometry analysis of HER2 and MDR1 expression in KPL-4 P and TR cells, and tumor derived KPL-4 TR cells. Supplemental Figure 3. qRT-PCR analysis of ErbB receptors and ligands in KPL-4 and BT-474M1 P and TR cells. Supplemental Figure 4. Flow cytometry analysis of HER2 expression in BT-474M1 P and TR cells. Supplemental Figure 5. Immunofluorescence detection of cell surface HER2 on BT-474M1 and T-DM1 resistant cells. Supplemental Figure 6. Analysis of T-DM1 uptake in BT-474M1 P and TR, and KPL-4 P and TR cells using radiolabeled T-DM1. Supplemental Figure 7. Treatment of KPL-4 TR cells with anti-IGF-1R antibody 10H5 or the selective c-met inhibitor PHA665725 does not restore sensitivity to T-DM1. Supplemental Figure 8. Upregulated EGFR does not mediate resistance to T-DM1 in KPL-4 TR cells. Supplemental Figure 9. Potential autocrine receptor-ligand interactions for T-DM1 resistance. Supplemental Figure 10. Comparable shRNA knockdown of PTEN in BT-474M1 cells in 2 different shRNA clones, #1 and #3. Supplemental Figure 11. Molecular alterations in BT-474M1 TR cells that did not confer T-DM1 resistance. Supplemental Figure 12. Expression of drug resistance transporters, assessed by qRT-PCR, in KPL-4 and BT-474M1 parental and TR cells. Supplemental Figure 13. The MDR1 inhibitor XR9051 and the BCRP inhibitor Ko143 do not affect cell viability Supplemental Figure 14. MRP4 siRNA in BT-474M1 TR does not reverse T-DM1 resistance. Supplemental Figure 15. MRP4 siRNA in KPL-4 TR does not reverse T-DM1 resistance. Supplemental Figure 16. Expression of SLC46A3 in KPL-4 parental and TR cells assessed by qRT-PCR. Supplemental Figure 17. SLC46A3 siRNA in BT-474M1 TR cells significantly reverses T-DM1 resistance.

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