Skip to main content
. 2020 Jun 17;39(30):5292–5306. doi: 10.1038/s41388-020-1360-y

Fig. 4. Reduced ECT2 levels in response to miR-1300 expression is associated with cytokinesis failure in U251 and KNS42 cell lines.

Fig. 4

a Confirmation of ECT2 knockdown at the protein level (by WB) following transfection with ECT2 siRNA smartpool. All experiments were performed in triplicate. The results were normalized to a scrambled-mimic control. b Transfection with ECT2 siRNA leads to an increase in binucleated cells. Binucleation phenotype scoring (3–6 fields of view (FOV) per condition, representing at least 100 cells) was performed on images taken on the Operetta imaging platform (×10 objective). c Immunofluorescence images comparing the actin and alpha-tubulin staining in U251 and KNS42 cells 72 h following transfection with 100 nM of either miR-1300 or ECT2 siRNA showing both the common effect on binucleation (DAPI, blue) and the comparative differences in actin (Phalloidin) and a-tubulin structures (Alexafluor488, green). Scale bar = 100 mm. d Ectopic expression of miR-1300 leads to decreased expression of ECT2 at the protein level. Statistical significance is expressed as follows: **p < 0.01; ****p < 0.0001. NB: Blot images were taken at 48 h time point since at 72 h, cells in control conditions have reached confluence, and were no longer expressing ECT2 as they are not dividing.