Fig. 3

ProT upregulates HOTAIR through NF-κB activation in bladder cancer cells. a–d ProT overexpression activates NF-κB and upregulates HOTAIR expression. J82 cells transduced with or without lentiviral vectors expressing ProT (J82/ProT) or vector control GFP (J82/GFP) were examined for ProT transcripts by RT-qPCR (a) and levels of ProT as well as NF-κB p65 and its phosphorylated form (p-NF-κB p65) by immunoblotting (b). c, d J82/ProT and J82/GFP cells were treated with JSH-23 (1 μM), TPCA-1 (1 μM), or the vehicle (DMSO) for 48 h, and levels of HOTAIR and ProT transcripts were assessed by RT-qPCR (c) and immunoblotting (d), respectively. e–g ProT knockdown reduces NF-κB activation and downregulates HOTAIR expression. TCCSUP cells transduced with or without lentiviral vectors expressing shRNA specific for human ProT (#17 and #19) or luciferase (Luc) were examined for ProT (n = 3–4) (e) and HOTAIR (g) transcripts by RT-qPCR as well as levels of ProT, NF-κB p65, and its phosphorylated form by immunoblotting (f). Representative immunoblots from three independent experiments and quantitative analysis of the indicated proteins are shown (b, f). h TCCSUP cells were treated with JSH-23 (1 μM), TPCA-1 (1 μM), or the vehicle (DMSO) for 48 h, and their HOTAIR levels were assessed by RT-qPCR. Values shown are mean ± SD (n = 3–4). The ratios of control cells were arbitrarily set to 100