Journal: Aging Cell

Article Title: Local Growth Hormone Facilitates Aging of the Colon Epithelial Microenvironment

doi: 10.1111/acel.70187

Figure Lengend Snippet: The npGH is expressed in senescent cells, and autocrine and paracrine GH promote β‐catenin nuclear translocation. (A‐B) Representative confocal images of npGH expression in senescent cells in (A) normal colon and (B) hyperplastic colon polyp. SA‐β‐gal, green; GH, red; colocalization, yellow. Scale bar = 100 μm. (C) Representative confocal image of hNCC expressing lentiGH. Arrow indicates GFP‐positive cell expressing GH (green) and active β‐catenin (red). Arrowhead indicates the cell cultured in proximity of a GH‐expressing cell also expressing intranuclear active β‐catenin. Scale bar = 20 μm. (D‐F) Representative cross‐sections of confocal Z‐stacks of hNCC showing localization of active β‐catenin (red) and nucleus (blue). (D) Active β‐catenin is outside the nucleus in GFP‐positive lentiV hNCC, (E) in the nucleus of GFP‐positive lentiGH hNCC, and (F) in the nucleus of GFP‐negative hNCC cocultured with lentiGH hNCC.

Article Snippet: Tissues were permeabilized in 1% Triton X100 for 30 min, followed by blocking in 10% goat serum for 1 h. Tissues were stained overnight at 4°C with anti β‐gal mouse monoclonal antibodies (dilution 1:50) and anti‐hGH rabbit antibodies (dilution 1:100), both from LSBio (cat#LS‐ B10989 and cat#LS‐B4199, respectively) followed by goat anti‐mouse Alexa 488 and goat anti‐rabbit Alexa 568 secondary antibodies, both from Invitrogen (cat#A32723 and #A11031, respectively).

Techniques: Translocation Assay, Expressing, Cell Culture