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. 2018 Feb 8;32(7):3583–3596. doi: 10.1096/fj.201701059R

Figure 4.

Figure 4

Inhibition of NAMPT expression induces SFTPB mRNA via transcriptional regulation in H441 cells. A) SFTPB mRNA levels were increased in H441 cells following siRNA knockdown of NAMPT. RNA was isolated from H441 cells transfected with control scrambled RNA and NAMPT siRNA for 48 h and subjected to quantitative RT-PCR. The relative quantification ± sd results represent 2 independent experiments, each analyzed in 5 technical replicates (n = 10). *P < 0.05. B) The effect of NAMPT expression on the SFTPB gene transcription. RNA capture was performed on H441 cells transfected with either control scrambled RNA or NAMPT siRNA for 48 h. The relative quantification ± sd of SFTPB mRNA levels was determined by quantitative RT-PCR analysis (n = 6 technical replicates). *P < 0.05. C). The effect of NAMPT expression on histone H3 hyperacetylation of the SFTPB promoter. An aliquot of the cells, cultured as in panel B, was harvested at 48 h, and the acetylation status of histone H3 in nucleosomes was assessed by a ChIP assay with an anti-acetylated histone H3 (K9) antibody and a primer pair specific for the SFTPB promoter. Input DNA was amplified as a control. D) Quantification of ChIP assays (n = 4). *P < 0.005.