Fig. 6

NF-κB-triggered α-Actinin-2 positive auto-regulatory loop enhances ACTN2 gene transcription. A and B α-Actinin-2, RelA, α-Actinin-2 siRNAs and/or RelA siRNAs were cotransfacted with (A) ACTN2 Luc-R or (B) 3xRelA/NF-κB Luc-R into SNU-16 cells for 12 h, then dual-luciferase reporter assays were performed. The statistical analysis information in A, B: The bars indicate the SD. The results are expressed as the mean ± SD of three independent experiments. *P < 0.05 using Ordinary one-way ANOVA. C NAGC and HAGC tissue sections obtained after infusion were immunostained for α-Actinin-2 (green) and nucleus (blue). D AGS cells were transfected with α-Actinin-2 for 24 h, then the PM, Cyto and Nu of cells were separated by the Subcellular Protein Fractionation Kit. IP assays were performed in PM, Cyto and Nu by using α-Actinin-2 antibodies. E and F α-Actinin-2, RelA and/or α-Actinin-2 siRNAs were cotransfacted into SUN-16 cells for 24 h, then cell nuclear extracts were subjected to ChIP analysis using the p-RelA antibody. E Western blotting analysis with the p-RelA antibody demonstrates the IP specificity and efficiency. F DNA isolated and purified from immunoprecipitated material was amplified with primers spanning the RelA binding site (Fs/Rs) or primers far away from the RelA binding site (Ff/Rf) of the ACTN2 gene promoter