Enhanced 4-Hydroxynonenal Resistance in KEAP1 Silenced Human Colon Cancer Cells
Figure 1
Effect of KEAP1 knockdown on NRF2 activity in HT29 cells. (a) The mRNA level for KEAP1 was determined by real-time PCR analysis for relative quantification in HT29 cells expressing scRNA (sc) and KEAP1-specific shRNA (shKEAP1). At the same time, the expression levels of KEAP1 were determined in HT29, sc, and shKEAP1 cells using conventional PCR analysis. (b) ARE-driven luciferase activity was monitored in HT29, sc, and shKEAP1 HT29 cells. (c) The nuclear level for NRF2 protein was determined in HT29, sc, and shKEAP1 HT29 cells. Lamin B levels were monitored as a loading control. (d) The basal mRNA levels for NQO1 and GCLC were determined in HT29, sc, and shKEAP1 HT29 cells. (e) Cellular total GSH contents were measured in the sc and shKEAP1 HT29 cells. The values are relative levels with respect to the sc group and are the means ± SD of 3-4 experiments. compared with the sc control.
