Figure 3. Relative levels of phosphorylated and total β-catenin at early stages of colonic crypt hyperplasia in relation to cellular CKIε levels.

A.Western blots showing relative levels of phosphorylated and total β-catenin in non-infected (N) and days 1–12 colonic crypt samples. β-Actin was used as loading control (n=5). B. Cellular levels of CKIε in non-infected (N) and Days 1–12 colonic crypt samples. +C is calyculinA-treated HEK293 cells used as positive control. C. Sub-cellular distribution of phosphorylated/total β-catenin in relation to relative levels of CKIε. Crypts isolated from uninfected (N ) and Days 6–34 days post-infected distal colons were fractionated into TritonX-100-soluble (C) and TritonX-100-insoluble (cytoskeletal, D) components and probed with antibodies to β-cat⁴⁵ or β-cat^33,37/41 and total β-catenin (β-cat). β-Actin was used as loading control. E. CKIε expression and co-immunoprecipitation (co-ip) with β-catenin during TMCH. Ei. Cellular crypt extracts prepared from normal uninfected (N) and Days 6–34 post-infected distal colons were probed with anti-CKIε. β-Actin was used as loading control (n=5). Eii. Co-ip with anti-CKIε in N and Days 6–27 colonic crypt extracts followed by blotting with anti-β-catenin and CKIε, respectively. F. Akt and PKA-catalyzed phosphorylation of β-catenin at Ser⁵⁵². Western blotting of crypt cellular extracts (N and Days 6–34) with an antibody recognizing Ser-552 in β-catenin. β-Actin was used as loading control (n=5).