Journal: bioRxiv
Article Title: Mechanically-induced Septin Networks Protect Nuclear Integrity
doi: 10.64898/2026.01.20.700414
Figure Lengend Snippet: A) SEPT2-Halo cells were treated with non-targeting (NT) or SEPT7 shRNAs. Whole-cell lysates were probed with SEPT7 (left) or SEPT2 (right) antibodies via Western blot. B) Fixed images of SEPT2-Halo NT (left) and SEPT7 (right) shRNA cells labeled with Halo dye and stained for actin (phalloidin). C) NT and SEPT7 shRNA cells were plated on a transwell with 3 µm pores and allowed to migrate overnight. Transwells were extracted, fixed, and immunostained for lamin A/C. D) Nuclear aspect ratio on top and bottom of the transwell was measured. Experimental replicates and total cells imaged for NT were 4 and top = 250, bottom = 54, for shRNA#1 were 4 and top =278, bottom= 59, and for shRNA#2 were 4 and top = 329, bottom = 77. A 2-way ANOVA was used to determine significance between top and bottom for each condition. E) NT and SEPT7 shRNA cells exogenously expressing 3xNLS-GFP were imaged unconfined and confined to 2.5 µ m. F) The percentage of nuclei that ruptured upon confinement was measured. Circles indicate replicate means and bars indicate mean ± SD. Experimental replicates and total cells imaged for NT were 5 and 474, for shRNA#1 were 5 and 310, and for shRNA#2 were 4 and 189. A one-way ANOVA was used to determine significance. For all statistics * = p < 0.05, and *** = p < 0.001.
Article Snippet: Anti SEPT7 (13818-1-AP; Protein Tech; WB 1:2000).
Techniques: Western Blot, shRNA, Labeling, Staining, Expressing