2.7. Statistical Analysis

To compare nuclear dimensions in different channel widths, a one-way ANOVA with Holm-Sidak’s multiple comparisons test, in the case of normally distributed data, or Kruskal–Wallis test with a Dunn’s post hoc test, in the case of non-normally distributed data, was completed. Data were pooled from at least three independent trials, except in 2D data for MSC and MDA-MB-231, which were pooled from two independent trials. A significance level of 0.05 was used. Error bars report the standard error of the mean. Select graphs were unable to fit all comparisons, in which case we refer readers to the Figure 1H and supplementary tables for full statistical analysis.

Quantification of 2D nuclear morphology in confinement. Images are shown for mesenchymal stem cells (MSCs) within microchannel widths of (A) 3 μm and B) 50 μm. Images are shown for L929 cells within microchannel widths of (C) 3 μm and (D) 50 μm. Images are shown for MDA-MB-231 cells within microchannel widths of (E) 3 μm and (F) 50 μm. In panels A–F cells were fixed and stained for actin (green) and the nucleus (blue). Color channels were altered individually for optimal visualization. Scale bar represents 10 μm in panels A–F. (G) Definition of the nuclear major and minor axes. Also shown are quantifications of the nucleus (H) area, (I) minor axis, and (J) major axis of MSCs, MDA-MB-231 cells, and L929 cells. Markers on line graphs report mean ± SEM of n cells, pooled from N ≥ 2 independent experiments with n(3 μm, MSC) = 9; n(3 μm, MDA-MB-231) = 178; n(3 μm, L929) = 26, n(6 μm, MSC) = 25; n(6 μm, MDA-MB-231) = 114; n(6 μm, L929) = 21; n(10 μm, MSC) = 21; n(10 μm, MDA-MB-231) = 98; n(10 μm, L929) = 80; n(20 μm, MSC) = 104; n(20 μm, MDA-MB-231) = 288; n(20 μm, L929) = 44; n(50 μm, MSC) = 289; n(50 μm, MDA-MB-231) = 620; n(50 μm, L929) = 136. Full statistical information for panels H–J is provided in Supplemental Tables S1–S3.