Figure 1.

HEK cultured on BMH exhibit increased cell density and decreased proliferation. (a) Representative bright-field images of HEK cultured on TCP and BMH, 0 and 24 h after treatment with 1.5 mM CaCl₂. Scale bars: 20 µm. (b) Quantification of cell density in a defined cell colony area of 10,000 µm² for each culture dish 0 and 24 h after CaCl₂ treatment (Solid columns = 0 h 1.5 mM CaCl₂, Striped columns = 24 h 1.5 mM CaCl₂). Data represent mean ±SEM, n = 3, statistical significance was assessed using one-way ANOVA with Tukey’s post hoc test, n.s. = non-significant (i.e., 50 kPa versus 8 kPa [low Ca⁺²]; 8 kPa versus 4 kPa [high Ca⁺²]), ** p ≤ 0.001. (c) Western blot of E-cadherin expression in HEK cultured on TCP and 4 kPa BMH following 1.5 mM CaCl₂ treatment. GAPDH levels indicate equal loading of proteins. (d) Representative confocal microscopy images of EdU proliferation assay on HEK cultured on TCP and BMH. DAPI stain denotes nuclei. Scale bars: 20 µm. (e) Quantification of the percentage of EdU positive cells grown on TCP and BMH cell culture dishes. Data represent mean ±SEM, n = 3, statistical significance was assessed using one-way ANOVA with Tukey’s post hoc test, * p ≤ 0.05, *** p ≤ 0.0001.