Fig. 5: HMGA1/p27/stathmin axis regulates migration in MDA-MB-231 cells.

A Lysates from MDA-MB-231 cells transfected with siCTRL and siHMGA1 were immunoprecipitated with stathmin and non-specific IgG antibodies. The amount of co-immunoprecipitated p27 is visualized by western blot analysis (the specific band is indicated by the arrow). The asterisk indicates the aspecific band due to the presence of immunoglobulines. Inputs are shown on the right. B p27-stathmin proximity was measured by in situ PLA in MDA-MB-231 cells transfected with siCTRL and siHMGA1. In situ PLA is indicated by green signals of the rolling cycle amplification products. Nuclei (blue) were counterstained using TO-PRO-3. Scale bar, 18 μM. Quantification of the number of fluorescent puncta per cell is shown on the right (n = 5 different fields). **P < 0.01; two-tailed Student’s t-test. C Quantification of wound-healing assay performed in MDA-MB-231 cells upon silencing of HMGA1 (siHMGA1), stathmin (siSTMN) and p27 (sip27). The results are presented as the mean of percentage of the wound closure relative to control (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001; two-tailed Student’s t-test. A representative western blot of HMGA1, stathmin and p27 expression after silencing is reported at the bottom of the graph. Representative pictures are reported on the bottom of the panel. D Kaplan–Meier survival curves of Relapse Free Survival (RFS) of 3951 breast cancer patients and Distant Metastasis Free Survival (DMFS) of 1746 BC patients investigated with Kaplan–Meier plotter (KM-plotter, https://kmplot.com/analysis/index.php?p=background) and containing information from GEO, EGA, and TCGA. The data investigated were based on the RNA expression of HMGA1/p27/STMN together (p27 is analyzed in inverted expression).