Figure 2

ER stress-mediated Lcn2 transcription is dependent on NF-κB activation in murine prostate cancer cells. (A) TC1.pNGL cells were treated with Tg (300 nM) or vehicle control (Veh), or LPS, for the indicated times. Cells were analyzed for EGFP reporter fluorescence by flow cytometry. Results shown are representative of 2-5 experiments. (B) TC1.pNGL cells were treated with Tg with or without 10 mM 4-phenylbutyric acid (PBA), or vehicle only. RNA was isolated and analyzed for UPR activation by RT-qPCR. Data columns indicate the fold difference in transcript level between drug- and vehicle-treated TC1.pNGL cells. Error bars represent SEM of 2-3 biological replicates representative of 3 independent experiments. (C) TC1.pNGL cells were treated as in (B) and analyzed for EGFP reporter fluorescence by flow cytometry. Cell viability of all treatment groups was > 99%. (D) TC1.pNGL cells from (B) were also analyzed for Lcn2, Il-6, and Il-23p19 transcription by RT-qPCR. Data columns indicate the fold difference in transcript level between drug- and vehicle-treated TC1.pNGL cells, and error bars represent SEM of 2-3 biological replicates representative of 3 independent experiments. Statistical analysis was performed using an unpaired two-tailed t test (**p < 0.01; ***p < 0.001). (E) Untreated TC1 cells (lane 1) or TC1 treated with vehicle control (lane 2), Tg (lane 3), or Tg + PBA (lane 4), as indicated above, were analyzed for Lipocalin 2 production by Western blot.