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. 2025 Aug 22;11(34):eadv6937. doi: 10.1126/sciadv.adv6937

Fig. 2. AKR1B10 curbs CRC metastasis in vitro and in vivo.

Fig. 2.

(A) Western blot analysis of AKR1B10 protein levels in human CRC (hCRC) cell lines. (B) Validation of AKR1B10 knockdown efficiency in HCT116 and SW1116 cells via Western blot and assessment of cell proliferation with Cell Counting Kit-8 (CCK-8) assays (n = 4). (C) Verification of AKR1B10 overexpression in LoVo cells by Western blot, with corresponding CCK-8 proliferation analysis (n = 4). (D and E) Representative images (D) and statistical analysis (E) of xenograft tumor volumes in nude mice implanted with HCT116 (shNC&shAKR1B10) cells (n = 5 mice per group). (F and G) Representative transwell assay images (left) and quantification (right) in HCT116/SW1116 cells (F) or LoVo cells after 48 hours (G). (H and I) Representative clonogenic assay images and quantification in HCT116/SW1116 cells (H) or LoVo cells (I). (J) H&E staining of lung metastatic nodules following intravenous injection of LoVo (Ctrl&AKR1B10-OE) cells. Ctrl, control; OE, overexpression. Scale bars, 250 μm. (K and L) Representative images (K) and quantification of lung tumor nodule area per section (L) in nude mice intravenously injected with HCT116 (shNC&shAKR1B10) cells (n = 5 mice per group). (M and N) Representative images and quantification of liver metastatic nodules in nude mice after splenic injection of HCT116 (shNC&shAKR1B10) cells (M), with corresponding H&E staining of liver metastatic lesions (N) (n = 5 mice per group). (O) mRNA expression levels of AKR1B10 in primary tumors (n = 56) and metastases (n = 27; liver = 23, peritoneum = 3, and lung = 1) based on GSE28702. (P and Q) Representative IHC images (P) and quantification (Q) of AKR1B10 in paired primary CRC tissues and liver metastatic lesions from 20 patients. Scale bars, 50 μm. (F to I) n = 3. Data are presented as mean ± SD, with two-way analysis of variance (ANOVA) test in (C) and (E) or unpaired Student’s t test (F to I, L, M, O, and Q). *P < 0.05, **P < 0.01, and ***P < 0.001.