10.1371/journal.pone.0088365.g005 Slobodan Culina Slobodan Culina François-Xavier Mauvais François-Xavier Mauvais Hsiang-Ting Hsu Hsiang-Ting Hsu Anne Burgevin Anne Burgevin Suzanne Guénette Suzanne Guénette Anna Moser Anna Moser Peter van Endert Peter van Endert HeLa Tet-on cells expressing H-2K<sup>b</sup> were nucleofected with 4×100 nM siRNA. Public Library of Science 2014 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 14:45:56 Figure https://plos.figshare.com/articles/figure/_HeLa_Tet_on_cells_expressing_H_2K_b_were_nucleofected_with_4_215_100_nM_siRNA_/928416 <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>