Normal human urothelial cells hucs

Polyclonal antibodies against cyclin E, CDK2, CDK6, cyclin D1, p53, p19ARF, p21WAF1, p27KIP1 and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-MMP-9 polyclonal antibody was obtained from Chemicon International (Billerica, MA, USA). miR-892b (5'-CACUGGCUCCUUUCUGGGUAGA-3') and miR-892b inhibitor were designed and synthesized by Genolution Pharmaceuticals, Inc. (Seoul, Korea).
Cell cultures. Human bladder carcinoma cell lines (EJ, 5637 and T24) were purchased from the American Type Culture Collection (ATCC; Manassas, vA, USA). The cells were maintained in Dulbecco's modified Eagle's medium (DMEM) with glucose supplemented with 10% fetal calf serum, L-glutamine and antibiotics (Biological Industries, Beit Haemek, Israel) at 37˚C in a 5% CO 2 humidified incubator. Normal human urothelial cells (HUCs) were purchase from ScienCell Research Laboratories (Carlsbad, CA, USA). The cells were maintained in urothelial cell medium with supplements according to the manufacturer's protocol.

Antibodies for ERK, phospho-ERK, p38, phospho-p38, JNK, and phospho-JNK were purchased from Cell Signaling Technology (Danvers, MA, uSA). Polyclonal antibodies against cyclin D1, cyclin E, CDK2, CDK6, p53, p21 CIP1/WAF1 , p27 KIP1 , GAPDH, Ets-1, Sp-1, ATF-2, and CREB were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, uSA). The anti-MMP-2 antibody was obtained from Chemicon international (Billerica, Ma, uSa). mir-106a (5'-AAAAGuGCuuACAG uGCAGGuAG-3') and mir-106a inhibitor were obtained from Genolution (Seoul, Korea).
Cell cultures. Human bladder carcinoma cell lines (EJ, 5637, and T24) were purchased from the American Type Culture Collection (aTCC, Manassas, Va, uSa). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum, l-glutamine, and antibiotics (Biological Industries, Beit Haemek, Israel) at 37˚C in a 5% CO 2 humidified incubator. Normal human urothelial cells (HuCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, uSA). The cells were grown in the medium specific for HuCs with supplements according to the manufacturer's protocol.