Figure 1
Knockdown of SETD7 represses NF-ĸB and the expression of proinflammatory cytokine under oxidative stress.
siRNA knockdown of SETD7 in Beas-2B cells was assessed by qRT-PCR (a) and western blot using antibody against SETD7 (b). *** indicates P < 0.001 (Student’s unpaired t test). (c–f) Transcriptional level of RELA, NFKBIA, IL-6 and IL-8 was measured by qRT-PCR after SETD7 siRNA transfection under unstimulated, CSE- or H2O2-stimulated conditions. Relative fold change of mRNA level was quantified by comparison with cells transfected with scramble siRNA without stimulation (mock). (g,h) IL-6 and IL-8 cytokine production under normal and ROS-stimulated condition was quantified by MSD. (i) Relative enrichment of H3K4me1 at the promoter region of RELA by qChIP. Beas-2B cells treated with scramble or siSETD7 siRNA were stimulated by H2O2 and their chromatin was fixed for the experiments. The level of H3K4me1 was shown by % of input of each individual sample. Data were expressed as mean ± standard error of the mean (SEM). Relative fold change was calculated by comparing each sample group with scramble siRNA-treated cells without stimulation. Two negative control sites locate at the gene desert were included for each experiment. n = 3. * indicates P < 0.05; ** indicates P < 0.01 (two-way ANOVA).
