FIGURE 3:

Nuclear area, cell area, and shape of cells revealed that all cell lines appeared morphologically different when cultured on 3 kPa and 40 kPa hydrogels. (A, B) All four cell lines had larger nuclei and cell area when cultured on 40 kPa hydrogels compared with 3 kPa hydrogels. (C) Analysis of form factor, representing the circulatory of cells (1 = a perfect circle) showed that MCF10A, 143B, and MDA-MB-231 cells appeared more circular when cultured on 3 kPa hydrogels. (D) Aspect ratio (X/Y) revealed that 143B cells had a larger X/Y ratio when cultured on stiff substrates. (E) Outlines of nuclear and cell images showing differences in size and morphology of cell cultured on both conditions. Outlines were obtained from images taken at 20× magnification and visualized in multicolor images by CellProfiler using F-actin for cytoplasmic and DAPI for nuclear boundary recognition. Scale bar = 100 µm. Data are shown as mean ± SEM. Numbers of cells used in analyses were indicated per bar graph. *, p < 0.05; **, p < 0.01; and ****, p < 0.0001.