Culina, Slobodan Mauvais, François-Xavier Hsu, Hsiang-Ting Burgevin, Anne Guénette, Suzanne Moser, Anna Endert, Peter van HeLa Tet-on cells expressing H-2K<sup>b</sup> were nucleofected with 4×100 nM siRNA. <p>Twenty-four hours later, the cells were electroporated with inducible plasmids encoding amyloid beta preceded by a signal peptide (+SP) or not (−SP), or encoding proinsulin, all tagged with the S8L peptide at the C-terminus. Protein expression was induced immediately by addition of 1 µg/ml doxycylin. Forty-eight hours later, K<sup>b</sup>/S8L complexes on the cell surface were detected using mAb 25D1.16 (Ab1) followed by FITC-labeled goat anti-mouse Abs (Ab2) and Alexa488-labeled goat anti-FITC Ab (Ab3). Control samples were HeLa-K<sup>b</sup> cells pulsed for 2 h with 10<sup>−8 </sup>M S8L and stained with Abs 1, 2 and 3 or with Abs 2 and 3 only, as well as peptide-pulsed HeLa cells expressing H-2K<sup>d</sup> stained with Abs 1, 2 and 3. One of two experiments is shown.</p> Anatomy and physiology;Immune physiology;Immune cells;Biochemistry;enzymes;Enzyme classes;hydrolases;metabolism;Protein metabolism;immunology;Antigen-presenting cells;Antigen processing and recognition;Immune response;Immune system;immunity;Major histocompatibility complex;Model organisms;Animal models;Animal types;Laboratory animals;tet-on;cells;expressing;nucleofected;nm 2014-02-07
    https://plos.figshare.com/articles/figure/_HeLa_Tet_on_cells_expressing_H_2K_b_were_nucleofected_with_4_215_100_nM_siRNA_/928416
10.1371/journal.pone.0088365.g005