Journal: Aging cell

Article Title: Amyloid-beta(1-42) alters structure and function of retinal pigmented epithelial cells.

doi: 10.1111/j.1474-9726.2009.00456.x

Figure Lengend Snippet: Fig. 5 Effects of OAβ(1-42) on cell attachment, tight junctions and permeability in ARPE-19 cells. (A) Effects of a 6-h pretreatment with OAβ(1-42) at the indicated concentrations on ARPE-19 cell attachment. Cell adhesion was measured by counting surface-attached cells at the indicated time after cell replating. The percentage of decrease of cell adhesion was compared to that in cells treated with PBS and with the inactive reverse control peptide Aβ(42-1). (B) F-actin structure was detected by rhodamine-labelled phalloidin and occludin immunochemistry was performed on ARPE-19 cells treated with OAβ(1-42) at 10 μM for 6 h as described in Experimental procedures. Arrows show the cell membrane staining for occludin. Without the anti-occludin antibody, cells treated with OAβ(1-42) or PBS and incubated with the secondary antibody coupled to FITC were not labelled (data not shown). Magnification, ×40. Scale bar = 50 μm (C) ARPE-19 cells were treated with or without OAβ(1-42) at 10 μM for 2 h. Total RNA was extracted and reverse-transcribed. The levels of claudin 1 (CLDN1), occludin (OCLN), Par-3 (PAR3) and zonula occludens-1 (ZO-1) expression were determined by quantitative PCR. (D) The effects of OAβ(1-42) on selective permeability were analysed in ARPE-19 cells treated with or without OAβ(1-42) at 10 μM for 2, 6 and 24 h and assessed with dextran-FITC with mean molecular weight of 40 kDa. Selective permeability in the inactive reverse control peptide treatment group did not differ from that in control cells treated with vehicle alone (data not shown). Similar results were obtained in three independent experiments. Data are the mean ± SD, *P ≤ 0.05.

Article Snippet: We used PBS/milk (1 × PBS, 15% milk) for saturation, and incubated cells with antibodies directed against acrolein (Novus Biologicals/Interchim) or occludin (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Cell Attachment Assay, Permeability, Control, Membrane, Staining, Incubation, Reverse Transcription, Expressing, Real-time Polymerase Chain Reaction, Molecular Weight

Journal: Frontiers in microbiology

Article Title: Early-Life Intervention Using Exogenous Fecal Microbiota Alleviates Gut Injury and Reduce Inflammation Caused by Weaning Stress in Piglets.

doi: 10.3389/fmicb.2021.671683

Figure Lengend Snippet: FIGURE 7 | Intestinal mucosal tight junction proteins expression of three groups. (A,B) Western blot measurements of the expression levels of ZO-1 and occludin in jejunum and colon. (C,D) The distributions of ZO-1 and occludin in jejunum and colon were determined by immunohistochemical staining (150×). (E,F) Western blot measurements of the expression levels of zonulin in jejunum and colon. The symbol “*” represented that the W and FW groups were compared with the S group, the symbol “#” represented that the FW group was compared with the W group. Data are shown as mean ± SEM for the three different experiments (*p < 0.05, **p < 0.01, #p < 0.05, and ##p < 0.01) (S, Sucking; W, Weaned; FW, FMT+Weaned).

Article Snippet: The following antibodies were used in the western blot assays: ZO1 (ThermoFisher 61-7300), occludin (Biorbyt orb11181), zonulin (Abcam ab231976).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining