Journal: bioRxiv

Article Title: Lamin A/C mediated invaginations in the nuclear surface allow the nucleus to pass unimpeded through a dense array of fiber-like obstacles

doi: 10.1101/2022.03.10.483838

Figure Lengend Snippet: (A) Schematic shows the calculation of a differential deflection vector as the vectorial change in deflection of the micropost after nuclear contact, and subsequent projection onto the nuclear surface normal. (B) A representative example of the calculated differential deflection vector (blue) and the corresponding normal component (white arrows) in a nuclear invagination (Scale bar is 5 μm). (C) Plot of the magnitude of the component of the differential vector along the surface normal against time for the example in (B). Time t = 0 refers to the time-point before the first nucleus-micropost contact. (D) Plot of the differential vector along the surface normal corresponding to 3C against the depth of indentation. (E) Plot shows the magnitude of force for mean values pooled from n = 10 cells from at least three independent experiments. Grey area represents SEM. The vertical dashed line (blue) indicates a plateau in the force. (F) Images show representative examples of fibroblasts on microposts (orange) treated with DMSO (control), Y-27 (25 μM), and Blebbistatin (50 μM) for 12 hours before fixation, followed by staining with Hoechst H33342 (blue, DNA) and Phalloidin (green, F-actin) (Scale is 10 μm in the top panel; Scale bar is 5 μm in bottom panel). (G) Plot of nuclear circularity of control and treated cells in (F), (“ns” p>0.05; Brown-Forsythe and Welch ANOVA test).

Article Snippet: The deflection of a micropost was calculated with an automated program written in MATLAB 2019a (MathWorks, Natick, MA, USA).

Techniques: Plasmid Preparation, Control, Staining