Journal: bioRxiv

Article Title: Mechanically-induced Septin Networks Protect Nuclear Integrity

doi: 10.64898/2026.01.20.700414

Figure Lengend Snippet: A) Cartoon depicting a septin hexamer and octamer with an approximation of our HaloTag and linker. B) Our SEPT2-Halo knock-in cells fixed and labeled with Halo dye (magenta) and immunostained for either SEPT2, SEPT7, or SEPT9 (green). Zoomed insets of the subnuclear region are shown to the right. C) Western blots of the parental MEF line and the SEPT2-Halo cells validating the knock-in did not change septin expression. The SEPT2-Halo band is shifted approximately 30 kDa larger, consistent with the approximate size of a HaloTag.

Article Snippet: Anti SEPT7 (13818-1-AP; Protein Tech; WB 1:2000).

Techniques: Knock-In, Labeling, Western Blot, Expressing

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