Fig. 1.

Lack of USF2 in cells affects morphology, proliferation and migration. (A) Representative Western blot analysis of USF2 protein levels in control (scrambled), USF2-deficient (ΔUSF2) and ΔUSF2 MEFs re-expressing a V5-tagged USF2 (ΔUSF2+USF2). (B) Light microscopy images of control, ΔUSF2 and ΔUSF2+USF2 MEFs stained with crystal violet. (C) Real-time proliferation rate of control, ΔUSF2 and ΔUSF2+USF2-MEFs. *significant difference, p ≤ 0.05. (D) Cell cycle distribution of control and ΔUSF2 cells. Histograms display cells in the G1 (blue fraction), S (yellow), and G2/M (green) phase of the cell cycle. (E) Quantification of cell cycle distribution. *significant difference control vs ΔUSF2 cells, p ≤ 0.05. (F, G) Apoptosis in control and ΔUSF2 MEFs was assessed by Annexin-V/P staining, measured by flow cytometry. (H) Real-time cell wound closure analysis of control, ΔUSF2 and ΔUSF2+USF2 MEFs; *significant difference, p ≤ 0.05. (I) Representative images of wound closure of control, ΔUSF2 and ΔUSF2+USF2 MEFs at 0 h, and 16 h after wound introduction. Scale bar 20 μm. (J) Migration was assessed by crystal violet staining and quantified by measuring the absorbance of crystal violet at 595 nm. The OD of ΔUSF2 cells was set to 100%. *significant difference control vs ΔUSF2 cells, **ΔUSF2 cells vs ΔUSF2+USF2 cells, p ≤ 0.05. (K) Representative images of the transwell migration assay. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)