Fig. 3.

Senescent fibroblasts significantly impair migrasomes formation A. mIHC of skin from young and aged mice. Skin sections from three 8-wFeek-old young mice and three 64-week-old aged mice were subjected to vimentin, TSPAN4, and WGA staining. Vimentin (green) identifies fibroblasts, while the colocalization of TSPAN4 (yellow) and WGA (red) denotes migrasomes. White arrows denote colocalization of Vimentin, TSPAN4 and WGA. Representative images are presented B. TEM of skin sample from young individuals. (a) Shows the dermal papilla structure; (b, c, d) present magnified views of migrasome-like structures. Yellow arrows denote migrasome-like structures, blue arrows indicate retraction fiber-like structures, and red arrows highlight potential migrasome precursor cells C. WGA staining images of young BJ cells and H₂O₂-induced senescent BJ cells, with magnified views D. SEM images of young BJ cells and H₂O₂-induced senescent BJ cells, with magnified views E. Quantification of migrasome numbers in young BJ cells and H₂O₂-induced senescent BJ cells using WGA staining. The experiment was independently repeated three times, with six random fields analyzed per group. Data are presented as mean ± SEM with significance (by normality and lognormality test followed by Mann-Whitney test) F. Quantification of migrasome numbers in young BJ cells and H₂O₂-induced senescent BJ cells was performed using scanning electron microscopy. The experiment was independently repeated three times, with three random fields analyzed per group. Data are presented as mean ± SEM with significance (by normality and lognormality test followed by Welch’s t test) G. Expression levels of migrasome markers in young BJ cells and H₂O₂-induced senescent BJ cells analyzed by Western blot