Figure 4.

NF-κB contributes to upregulation of CD166. (A) Promoter activities from CD166 NF-κB reporter in HepG2 cells were measured after overexpression of NF-κB components, as indicated, with FBS condition, *P < 0.05 and **P < 0.01 versus ‘Empty’ or knockdown either P65 or P50 using specific siRNA under SD, *P < 0.05, versus ‘NC’. (B) Effects of SD with or without BAY 11–7082 (100 μM) treatment on the amount and phosphorylation of IκBα protein. β-actin was treated as internal control. (C) Total RNA from cells was extracted after 12 h treatment with either siRNA or pcDNA3.1, as indicated. Then CD166 mRNA levels were measured using real-time PCR. **P < 0.01 level using t-test. (D) CD166 protein levels from cells under similar treatment as described in (C) were assayed by western blotting. (E) Effects of LPS or TNFα on CD166 expression before or after SD. (F) Open chromatin accessibility status of HepG2 cells under different treatment as indicated was measured via chart-PCR (29 reaction cycles) using either DNase I, Mae III or Sma I by same primer sets used in Figure 3F.