WT 64 NCI-H520 Non-small cell lung PRN1371 cancer WT Reduced mRNA 68 ZR-75-30 Breast, metastatic-ascites, invasive ductal carcinoma WT WT 77

ZR-75-1 Breast, metastatic-ascites, invasive ductal carcinoma WT WT 80 Huh7 Hepatocellular carcinoma WT Mut 84 BT474 Breast, primary, invasive ductal carcinoma WT Mut 86 GSK126 manufacturer PLC/PRF/5 Hepatocellular carcinoma WT Inactivated 92 Hep3B Hepatocellular carcinoma No Deletion 96 Low sensitivity (100 nM < GI50 < 1 μM) U2OS Osteosarcoma Less active WT 139 Hs578T Breast, metastatic, invasive ductal carcinoma WT Mut 143 MV4-11 Acute myeloid leukemia WT Mut 231 RS4;11 Acute myeloid leukemia WT Mut 254 HepG2 Hepatocellular carcinoma WT WT 273 MOLM-13 Acute myeloid leukemia WT Mut 315 Resistant (GI50 > 1 μM) A549 Non-small cell lung cancer WT WT >10 μM HCC1954 Breast, invasive ductal carcinoma Mut WT >10 μM

MDA-MB-361 Breast, metastatic-brain, adenocarcinoma WT No >10 μM MOLT-4 Acute lymphoblastic leukemia WT WT >30 μM N87 Gastric cancer WT WT >30 μM *WT, wild type; Mut, mutated. To determine the activity of TAI-1 in multidrug resistant (MDR) cell lines, established MDR cell lines were tested. MES-SA/Dx5 and NCI-ADR-RES are resistant to doxorubicin and paclitaxel, Seliciclib ic50 while Fluorometholone Acetate K562R cells are resistant to imatinib. TAI-1 was active in these cell lines showing nM GI50 (Table 2). Table 2 GI 50 s of TAI-1 and

commerically available drugs in cell lines   Cell line TAI-1 GI50(nM) Drug resistant cancer cell lines MEX-SA/Dx5 35 NCI/ADR-RES 29 K562R 30 Normal cell lines WI-38 >10 μM RPTEC >10 μM HuVEC > 9 μM HAoSMC > 9 μM *N.D, not determined. TAI-1 targets the Hec1-Nek2 pathway and induces apoptotic cell death To confirm the mechanism of action of TAI-1, we used established methods to evaluate the interaction of Hec1 and Nek2 and the consequences of disruption of interaction of the proteins [3]. Co-immunoprecipitation study shows that TAI-1 disrupted the binding of Nek2 to Hec1 in TAI-1-treated cells (Figure 2A). Disruption of Nek2 binding to Hec1 was shown to lead to degradation of Nek2 [3], and this was also confirmed for TAI-1 (Figure 2B). In addition, previous study also show that disruption of Hec1-Nek2 interaction leads to misaligned chromosomes.