Figure 5. NEK2 silencing phenocopies the loss of LIN9, inducing mitotic progression errors and restoring paclitaxel sensitivity.

MDA-MB-231 sensitive/parental and resistant cells were transfected with siNS, siLIN9, or siNEK2 for 3 days before staining with DAPI, Texas Red-X phalloidin, and γ-tubulin. A. RT-qPCR analysis to confirm NEK2 silencing, *=p<0.05 siNEK2 compared to control. The percent of cells with B. 3+ centrosomes, C. micronuclei, D. multinucleation, and E. dysmorphic nuclei were counted (minimum 150 cells per experiment). Graphs B-E are derived from the same experiment as Figure S2 A–D, but with siNEK2 data added. Distinct letters above bars indicate significant differences between groups(p<0.05). F and G. Kaplan-Meier curves of taxane and anthracycline-treated TNBC patients stratified by high or low NEK2 expression (quartiles) who experienced F. a pathological complete response (pCR) or G. residual disease (RD). H. MDA-MB-231 sensitive/parental and resistant cells were transfected with siNS or siNEK2 and then treated with increasing concentrations of paclitaxel for 4 days. Live cells were counted using trypan blue exclusion, *p<0.05 comparing MDA-MB-231 resistant siNS to siNEK2, and #p<0.05 comparing MDA-MB-231 sensitive/parental siNS to siNEK2. Panels A-E and H were repeated in 3 independent experiments in triplicate and are represented as means ± SD.