(A) Immunoblots of HT29 cell extracts without (control) or with CRISPR knockdown of ZNF277 expression. β-Actin was used as a loading control. (B) ZNF277 deficiency attenuates HT29 cell proliferation in vitro from 7 separate experiments. (C) ZNF277 deficiency attenuates xenograft growth. Time-course reveals reduced volume of ZNF277 CRISPR HT29 cell–derived xenografts (n = 8) compared with control xenografts (n = 7). (D) Representative images of s.c. and excised xenografts from HT29 ZNF277 CRISPR versus control cells. Arrows and arrowheads indicate control and HT29 ZNF277 CRISPR xenografts, respectively. (E) Reduced weights of xenografts with ZNF277 deficiency (n = 8). *P < 0.01 versus controls (n = 7); 2-tailed Student’s t test. (F) Representative microscopic images of control and ZNF277-deficient xenografts stained for H&E, p21WAF1, and p53. (G) ZNF277, p21WAF1, p53, and β-catenin immunoblots of proteins extracted from ZNF277 CRISPR cell– and control cell–derived tumors (2 separate tumors from each group). Values represent mean ± SD. Scale bar: 100 μM.