Figure 4. Nox4 interaction with prelamin in nuclei of AFSC. (A) Representative images of AFSC group I, II and III (faster to slower senescent cells) showing green signals of Prelamin A (Prelam A), and, in doubled magnification image in white square of group II, farnesylated Prelamin A (farn Prel). Scale bar= 10 µm. (B) Representative images of AFSC group II labelled with DAPI (blue), Nox4 (green) and Prelamin A (red) or farnesylated Prelamin A (red). Scale bar=10 µm. (C) Western Blot analysis revealed with anti-Prelamin A or the specific anti-Lamin A/C antibodies of total hAFSCs lysates of group II at passage 1, 4 and 8 and of group I, II and III at passage 4. Tubulin detection was performed as a loading control. Mevinolin (Mev) treatment was performed to show a prelamin A positive control. Data are representative of three independent experiments. (D) Western blot analysis of total lysate (TL) and immunoprecipitation (IP) experiment of TL with Prelamin A or farnesylated Prelamin A antibody then revealed with anti-Nox4. Preclearing samples were also loaded, in order to exclude non-specific signals. Data are representative of three independent experiments.(E) Western blot analysis of nuclear lysate (NL) and immunoprecipitation experiment of Nuclear lysate (IP Nu) and Cytoplasmic lysate (IP Cy) with Nox4 antibody then revealed with anti-Prelamin A. Mevinolin (Mev) treatment was performed to show a prelamin A positive control. Data are representative of three independent experiments.