Fig. 3.

ARID2 suppressed EMT and Snail transcription of HCC cells. (A and B) The expression of EMT associated factors in ARID2 knockdown and overexpressing HCC cells were examined by Western blot. (C and D) mRNA levels of Snail and E-cadherin in stably transfected HCC cell lines were examined by RT-qPCR. (E) Based on previous ChIP-seq data from HepG2 cells, ARID2 potentially bound at the promoter region of Snail gene. (F) Effect of ARID2 on activity of Snail promoter in stably transfected HCC cells was assessed by reporter assay. (G and H) Snail expression in metastatic lesions of intrahepatic metastasis mouse model and HCC mouse models was evaluated by immunohistochemistry. (Scale bar, 25 μm.) (I) The converse ARID2 and Snail expression pattern in clinical HCC tissues were assessed by immunohistochemistry. (J), (K) The correlation of expression between ARID2 and Snail at mRNA (n = 34, P = 0.0046) and protein (n = 287, P < 0.001) levels were analyzed. (L) A survival time comparison between HCC patients with ARID2^highSnail^low expression with patients with ARID2^lowSnail^high expression was performed, P = 0.0041. According to the immunostaining intensity of ARID2 and Snail (high, H-score ≥ 38; low, H-score < 38) protein, patients (shown in Fig. 1 E and F) were divided into ARID2^highSnail^high (n = 25), ARID2^highSnail^low (n = 59), ARID2^lowSnail^high (n = 181) and ARID2^lowSnail^low (n = 22) group. Data were analyzed using Student’s t test. All *P < 0.05, **P < 0.01, ***P < 0.001.