Journal: Materials Today Bio

Article Title: Low-toxicity vitrification of small-diameter arterial grafts by achieving a Cryo-Stasis state

doi: 10.1016/j.mtbio.2026.103439

Figure Lengend Snippet: CTX-Rads stabilizes the nuclear envelope via Lamin B1 to inhibit cell death. (A) Immunofluorescence staining of Lamin B1 in HUVECs after exposure to CPAs for the indicated durations. Cells were fixed with 4% paraformaldehyde without unloading. (B) Lamin B1 mean fluorescence intensity was quantified from (A) ( n ≥ 30 cells per group). (C) HUVECs were transfected with Lamin B1 overexpression plasmid (OE) or empty vector (Control) (n = 3). For (C – E), transfected cells were subjected to VS55 loading (exposure for 15 min) followed by one-step unloading. DNA damage was assessed by γ -H2AX immunofluorescence. Representative images are shown in panel C, and the percentage of γ -H2AX positive cells is quantified in (D). UT, untreated. (E) Cell viability of Lamin B1-overexpressing HUVECs was measured by CCK-8 assay (n = 3). (F, G) HUVECs were subjected to Lamin B1 knockdown. These cells were then exposed to CTX-Rads loading (exposure for 60 min or 90 min) followed by one-step unloading. DNA damage was evaluated in (F) and cell viability in (G) (n = 3). (H) CTX-Rads upregulates Lamin B1 to stabilize the nucleus and enhance cell survival during unloading, whereas VS55 compromises this protective mechanism. All experiments was performed in triplicate. Data: mean ± SD; Tukey’s test: ns, P > 0.05 ; * P ≤ 0.05 , ** P ≤ 0.01 , *** P ≤ 0.001 .

Article Snippet: They were then incubated with a rabbit anti-lamin B1 primary antibody (ABclonal, A1910; 1:200) followed by an Alexa FluorTM 488-conjugated goat anti-rabbit IgG secondary antibody (Thermo Fisher, Cat# A11034; 1:500).

Techniques: Immunofluorescence, Staining, Fluorescence, Transfection, Over Expression, Plasmid Preparation, Control, CCK-8 Assay, Knockdown