Rhodamine red x donkey anti mouse

FFPE sections were processed and stained as previously described (20 (link)). Images were acquired and processed as described above for histopathological analysis. The following primary antibodies were used: rabbit polyclonal anti‐C. rodentium antibody (1:50) (Statens Serum Institute, Copenhagen, Denmark), mouse anti-PCNA antibody (1:500) (catalog number ab29; Abcam), and rabbit anti-CHGB antibody (1:50) (catalog number 14968-1-AP; Proteintech). The following secondary antibodies were used: Alexa Fluor 555 anti-rabbit and rhodamine Red-X donkey anti-mouse (both 1:100) (Jackson Immunoresearch). DNA was stained with Hoechst 33342 or 4′,6-diamidino-2-phenylindole (DAPI) (1:1,000). PCNA measurements were obtained from at least 10 well-oriented crypts per mouse. Colonic sections from which fewer than 10 well-orientated crypts were observed were excluded from all analyses. The endocrine cell density (cells per square millimeter of crypt area) for each mouse was calculated by dividing the total number of CHGB-positive cells within the crypt area by the total crypt area. The total crypt area was calculated by tracing around the edges of colonic crypts and the lumen of the gut using the Contour(polygon) tool in Zen 2.3 (Blue version; Carl Zeiss MicroImaging GmbH, Germany).