fig8

Figure 8. Analysis of the impact of ANKRD6 knockdown on protein expression and related gene expression in Colon Adenocarcinoma cells. (A) Immunohistochemistry revealed cytoplasmic enrichment of ANKRD6 in COAD samples, with positive regions visualized as brown staining, whereas adjacent normal tissues exhibited weaker signals. (B) qPCR results showing that ANKRD6 mRNA levels were significantly reduced in siANKRD6-1, siANKRD6-2, and siANKRD6-3 transfected groups compared to the negative control (NC group) (P < 0.01), with knockdown efficiencies of 28%, 54%, and 33%, respectively. (C) qPCR results showing that mRNA levels of JNK, RHOA, and WNT7A in the siANKRD6-transfected groups were significantly lower than those in the control group (siCtrl) (P < 0.01). (D) Western blot analysis of WNT7A, JNK, and RhoA protein levels in siCtrl and siANKRD6 COAD cells, with GAPDH as the loading control. The right panels show densitometric quantification of protein bands normalized to GAPDH. WNT7A expression was significantly reduced after ANKRD6 knockdown (P < 0.05), whereas JNK and RHOA levels exhibited no significant change (ns). (ns: Not significant; *P < 0.05; **P < 0.01).