Journal: Aging Cell

Article Title: Environmental Enrofloxacin Exposure as a Modifiable Driver of Mitochondria‐Mediated Intestinal Aging and Barrier Dysfunction

doi: 10.1111/acel.70526

Figure Lengend Snippet: Effects of ENR exposure on intestinal barrier integrity in zebrafish. (A) Experimental design illustrating the exposure protocol. (B, C) Representative immunofluorescence images of Cdkn1a and quantification. (D, E) Representative immunofluorescence images of Cdkn2a and quantification. (F, G) Zebrafish intestinal permeability assessment using Smurf assay ( n = 10). (H) Hematoxylin and eosin staining of zebrafish intestinal tissues. (I) Quantification of goblet cells in zebrafish intestinal tissues. (J, K) Representative Periodic Acid‐Schiff (PAS) staining and quantification of mucus production in zebrafish intestinal tissue. (L–R) Representative immunofluorescence images of intestinal tight junction proteins (Occludin, Mucin‐2, Zo‐1, Claudin) and quantification. (S, T) Representative immunofluorescence images of intestinal CD3‐positive T cells and quantification. Data are presented as the mean ± standard error of the mean. Statistical significance was assessed using Student's t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Intestinal tissues were fixed (4% paraformaldehyde, 15 min), blocked (10% goat serum albumin, 0.4% Triton X‐100), and incubated with primary antibodies from Servicebio: Cdkn1a (GB11153, 1:300), Cdkn2a ( GB151143 , 1:300), Tomm20 ( GB151481 , 1:1000), Grp75 ( GB112273 , 1:650), Cox5a ( GB111676 , 1:500), Hsp60 ( GB112464 , 1:800), Cox4 (GB11250, 1:200), CD3 (GB13014, 1:100), Mucin‐2 (GB11344, 1:500), Occludin ( GB111401 , 1:750), Zo‐1 ( GB115686 , 1:1000), and Claudin‐1 ( GB112543 , 1:1000).

Techniques: Immunofluorescence, Permeability, Staining

Journal: Aging Cell

Article Title: Environmental Enrofloxacin Exposure as a Modifiable Driver of Mitochondria‐Mediated Intestinal Aging and Barrier Dysfunction

doi: 10.1111/acel.70526

Figure Lengend Snippet: Effects of fecal microbiota transplantation (FMT) on intestinal barrier function. (A) Experimental overview illustrating the procedure of FMT following antibiotic (ABX) treatment and ENR exposure. (B, C) Representative immunofluorescence images of intestinal CD3‐positive T cells and quantification after FMT. (D, E) Representative immunofluorescence images of hypoxia markers and quantification after FMT. (F–L) Representative immunofluorescence images of intestinal tight junction proteins (Mucin‐2, Occludin, Zo‐1, Claudin) and quantification after FMT. Data are presented as the mean ± standard error of the mean. Statistical significance was assessed using Student's t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Intestinal tissues were fixed (4% paraformaldehyde, 15 min), blocked (10% goat serum albumin, 0.4% Triton X‐100), and incubated with primary antibodies from Servicebio: Cdkn1a (GB11153, 1:300), Cdkn2a ( GB151143 , 1:300), Tomm20 ( GB151481 , 1:1000), Grp75 ( GB112273 , 1:650), Cox5a ( GB111676 , 1:500), Hsp60 ( GB112464 , 1:800), Cox4 (GB11250, 1:200), CD3 (GB13014, 1:100), Mucin‐2 (GB11344, 1:500), Occludin ( GB111401 , 1:750), Zo‐1 ( GB115686 , 1:1000), and Claudin‐1 ( GB112543 , 1:1000).

Techniques: Transplantation Assay, Immunofluorescence

Journal: Aging Cell

Article Title: Environmental Enrofloxacin Exposure as a Modifiable Driver of Mitochondria‐Mediated Intestinal Aging and Barrier Dysfunction

doi: 10.1111/acel.70526

Figure Lengend Snippet: Effects of PQQ treatment on ENR‐induced intestinal barrier damage. (A, B) Representative immunofluorescence images of Cdkn1a and quantification. (C, D) Representative immunofluorescence images of Cdkn2a and quantification. (E, F) Periodic Acid–Schiff (PAS) staining of zebrafish intestinal tissues and quantitative analysis. (G, H) Zebrafish intestinal barrier integrity assessed using the Smurf assay. (I–O) Representative immunofluorescence images of intestinal tight junction proteins (Mucin‐2, Occludin, ZO‐1, Claudin) and quantification. (P, Q) Western blot analysis and quantification of intestinal TNF‐α protein levels. Data are presented as the mean ± standard error of the mean. Statistical significance among the Control, ENR, and ENR + PQQ groups was assessed using one‐way ANOVA followed by Tukey's multiple‐comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Intestinal tissues were fixed (4% paraformaldehyde, 15 min), blocked (10% goat serum albumin, 0.4% Triton X‐100), and incubated with primary antibodies from Servicebio: Cdkn1a (GB11153, 1:300), Cdkn2a ( GB151143 , 1:300), Tomm20 ( GB151481 , 1:1000), Grp75 ( GB112273 , 1:650), Cox5a ( GB111676 , 1:500), Hsp60 ( GB112464 , 1:800), Cox4 (GB11250, 1:200), CD3 (GB13014, 1:100), Mucin‐2 (GB11344, 1:500), Occludin ( GB111401 , 1:750), Zo‐1 ( GB115686 , 1:1000), and Claudin‐1 ( GB112543 , 1:1000).

Techniques: Immunofluorescence, Staining, Western Blot, Control