Fig. 1

FKBPL overexpression induces differentiation of CSC-like colonies and downregulates DLL4 expression in MDA-MB-231 cells. A reduction in the number of holoclones (a) and a concomitant increase in the number of meroclones and paraclones (b) was observed, using MDA-MB-231 cells with stable FKBPL overexpression (A3), vs. MDA-MB-231 controls. 50 or 100 cells/cm² per well were seeded in a six-well plate containing DMEM + 10% FCS medium and incubated for 10 days at 37°C in a humidified atmosphere of 95% O₂/5% CO₂ before colonies were counted manually. (representative images in the inset; 1 – holoclones; 2 – meroclones; 3- paraclones). c Western blot of cell lysates collected from A3 or MDA-231 cells were probed for DLL4, FKBPL and GAPDH (representative blot in inset). Protein expression was quantified using ImageJ, adjusted to GAPDH and normalised to control. d Real-time qPCR analysis of DLL4 mRNA levels in MDA-MB-231 vs. A3 cells. The difference in gene expression was presented as a fold change relative to the expression of the housekeeping genes, GAPDH and ß –Actin. Data points are mean ± SEM. n ≥ 3. * p < 0.05, ** p < 0.01, ***p < 0.01 (t-test)