Anti cd3

Deeply anesthetized mice were transcardially perfused using 4% paraformaldehyde in phosphate-buffered saline (PBS). Gut tissue was removed and post-fixed overnight in 4% paraformaldehyde at 4 ℃ and equilibrated in 30% sucrose. Frozen 20 μm coronal colon sections were hydrated, immersed in 3% hydrogen peroxide in methanol, and incubated for 1 h with 5% normal goat serum at RT. Next, in a humidified chamber, mouse anti-Ly6G (1:200, Abcam), anti-CD3 (1:200, Novusbio), and anti-F4/80 (1:200, Proteintech) antibodies were incubated with sections overnight at 4 ℃. After washing in PBS, Ly6G, CD3, and F4/80IHC localization were determined using a HRP/DAB kit (Catalog number: 20774 and 20775, MilliporeSigma). Images were captured as described. We quantified immunostaining using ImageJ (NIH). For all sections, the same threshold settings and image exposure times were used.

The protein phosphatase inhibitor cocktail (P5726) and protease inhibitor cocktails (8340) were purchased from MilliporeSigma (Berlington, MA, USA). All antibodies are listed here; anti-integrin β3 (Abcam, Cambridge, UK, ab179473), anti-Aβ_1-16 (6E10) (Biolegend, San Diego, CA, USA) (803015), anti-c-Myc (Santa Cruz Biotechnology, Dallas, TX, USA) (sc-40), anti-β-galactosidase (Proteintech, Rosemont, IL, USA) (15518-1-AP), anti-actin (MilliporeSigma, Burlington, MA, USA) (A1978), anti-pan-cytokeratin (Thermo Fisher Scientific, Waltham, MA, USA, 53-9003-80), anti-Ki67 (Thermo Fisher Scientific, 14-5698-80), anti-Ly6G (Abcam, ab238132), anti-CD3 (Novusbio, Englewood, CO, USA) (NBP2-25186SS), and F4/80 (Proteintech, 28463-1-AP). The senescence β-galactosidase staining kit came from Cell Signaling (Danvers, MA, USA) (9860). Nuclear-fast red aluminum sulfate (0.1%) (1001210500), Alcian blue (1016470500), and hematoxylin and eosin (H&E) stains (HT110216) were supplied by MilliporeSigma.