Epithelium-Specific Ets-Like Transcription Factor 1, ESE-1, Regulates ICAM-1 Expression in Cultured Lung Epithelial Cell Lines

Figure 2

Effects of ESE-1 knockdown in A549 cells on the expression of ICAM-1. (a) Schematic diagrams of the helper-dependent adenovirus vectors (HD-Ad) that were used for the ESE-1 knockdown experiments. ESE-1-RNAi expresses two shRNAs from the murine U6 gene promoter, and C4HSU is used as an empty vector control which does not express any transgene. ITR: inverted terminal repeat; Ψ: packing signal. (b, c) ESE-1 and ICAM-1 mRNA expression in A549 cells 4 and 5 days after transduction with ESE-1-RNAi vector compared to that of C4HSU. Both groups were stimulated with TNF-α and IL-1β at 10 ng/mL each for 2 hours before cell lysis. The mRNA expression levels were normalized to GAPDH and the values were presented in as the mean ± SD, , . Statistics was performed as described in Materials and Methods. (d) A representative western blot analysis of ICAM-1 expression with (+) and without (−) ESE-1-RNAi, compared to that of the C4HSU vector control group on day 4 and day 5 after transduction. Since ICAM-1 levels were very low, subgroup of cells were stimulated with TNF-α and IL-1β (10 ng/mL each) for 16 hours to visualize ICMA-1 protein expression before cell lysis.

(a)

(b)

(c)

(d)