Research Paper Volume 15, Issue 15 pp 7513—7532

Figure 6. NORAD knockdown increased the binding of PGC-1α and SIRT1. ARPE-19 cell lines were transfected with siNORAD or siNC, after 96 hours, the interaction of NORAD with PGC-1α and SIRT1 was detected. (A) RIP assay confirmed the interaction of NORAD and PGC-1α. All groups of ARPE-9 were incubated with the anti-PGC-1α or anti-IgG antibody, and the precipitated complexes were analyzed with RT-qPCR to investigate the expression of NORAD. n = 3, ***P < 0.001. (B) RIP assay confirmed the interaction of NORAD and SIRT1. All groups of ARPE-9 were incubated with the anti-SIRT1 or anti-IgG antibody, and the precipitated complexes were analyzed with RT-qPCR to investigate the expression of NORAD. n = 3, ***P < 0.001. (C) ARPE-19 was transfected with siSIRT1 or normal contrast siNC. SIRT1 levels were analyzed through western blot. n = 3, ***P < 0.001 vs. siNC. (D) Immunoprecipitation and western blot analysis of acetyl-PGC-1α levels in ARPE-19 transfected with siNORAD, siSIRT1, or normal contrast siNC. (E) IP experiment confirmed the interaction of SIRT1 and PGC-1α. All groups of ARPE-9 were incubated with the anti-PGC-1α or anti-IgG antibody, and the precipitated complexes were analyzed with anti-SIRT1 by western blot. n = 3. (F) IP experiment confirmed the interaction of PGC-1α and SIRT1. All groups of ARPE-9 were incubated with the anti-SIRT1 or anti-IgG antibody, and the precipitated complexes were analyzed with anti-PGC-1α by western blot. n = 3.